An evaluation of reproductive toxicity studies and data interpretation of N-methylpyrrolidone for risk assessment: An expert panel review.

An expert panel was assembled to evaluate reproductive toxicology study data and their application to health risk assessment to provide input on the data quality, interpretation, and application of data from three multi-generation reproductive toxicity studies of N-methylpyrrolidone (NMP). Panelists were engaged using a double-blinded, modified Delphi format that consisted of three rounds. Key studies were scored using the U.S. Environmental Protection Agency's (EPA) questions and general considerations to guide the evaluation of experimental animal studies for systematic review. The primary conclusions of the panel are that one of the studies (Exxon, 1991) is not a high-quality study due to several design flaws that includes: (1) exceedance of the maximum tolerable dose in the high dose group; (2) failure to adjust feed concentrations of NMP during the lactation period, resulting in NMP doses that were 2- to 3-fold higher than nominal levels; and/or (3) underlying reproductive performance problems in the strain of rats used. For these reasons, the panel recommended that this study should not be considered for quantitative risk assessment of NMP. Exclusion of this study, and its corresponding data for male fertility and female fecundity, from the quantitative risk assessment results in a change in the identification of the most sensitive endpoint. Instead, changes in rat fetal/pup body weight, an endpoint previously selected by EPA, was identified as an appropriate basis for human health risk assessment based on a consideration of the best available science and weight of scientific evidence supported by the NMP toxicity database.

Grouping of PFAS for human health risk assessment: Findings from an independent panel of experts.

An expert panel was convened to provide insight and guidance on per- and polyfluoroalkyl substances (PFAS) grouping for the purposes of protecting human health from drinking water exposures, and how risks to PFAS mixtures should be assessed. These questions were addressed through multiple rounds of blind, independent responses to charge questions, and review and comments on co-panelists responses. The experts agreed that the lack of consistent interpretations of human health risk for well-studied PFAS and the lack of information for the vast majority of PFAS present significant challenges for any mixtures risk assessment approach. Most experts agreed that "all PFAS" should not be grouped together, persistence alone is not sufficient for grouping PFAS for the purposes of assessing human health risk, and that the definition of appropriate subgroups can only be defined on a case-by-case manner. Most panelists agreed that it is inappropriate to assume equal toxicity/potency across the diverse class of PFAS. A tiered approach combining multiple lines of evidence was presented as a possible viable means for addressing PFAS that lack analytical and/or toxicological studies. Most PFAS risk assessments will need to employ assumptions that are more likely to overestimate risk than to underestimate risk, given the choice of assumptions regarding dose-response model, uncertainty factors, and exposure information.

New developments in the evolution and application of the WHO/IPCS framework on mode of action/species concordance analysis.

The World Health Organization/International Programme on Chemical Safety mode of action/human relevance framework has been updated to reflect the experience acquired in its application and extend its utility to emerging areas in toxicity testing and non-testing methods. The underlying principles have not changed, but the framework's scope has been extended to enable integration of information at different levels of biological organization and reflect evolving experience in a much broader range of potential applications. Mode of action/species concordance analysis can also inform hypothesis-based data generation and research priorities in support of risk assessment. The modified framework is incorporated within a roadmap, with feedback loops encouraging continuous refinement of fit-for-purpose testing strategies and risk assessment. Important in this construct is consideration of dose-response relationships and species concordance analysis in weight of evidence. The modified Bradford Hill considerations have been updated and additionally articulated to reflect increasing experience in application for cases where the toxicological outcome of chemical exposure is known. The modified framework can be used as originally intended, where the toxicological effects of chemical exposure are known, or in hypothesizing effects resulting from chemical exposure, using information on putative key events in established modes of action from appropriate in vitro or in silico systems and other lines of evidence. This modified mode of action framework and accompanying roadmap and case examples are expected to contribute to improving transparency in explicitly addressing weight of evidence considerations in mode of action/species concordance analysis based on both conventional data sources and evolving methods.

Field efficacy and safety of an oral formulation of the novel combination anthelmintic, derquantel-abamectin, in sheep in New Zealand.

AIM: To evaluate the efficacy and safety of the novel anthelmintic combination, derquantel-abamectin, against gastrointestinal nematode populations in sheep, under field-use conditions. METHODS: Controlled faecal egg count reduction tests (FECRT) were conducted in New Zealand in 14 trials, covering a range of geographic locations, farming enterprises, breeds, nematode populations, and anthelmintic-resistance profiles. Enrolled animals were naturally infected with mixed populations of gastrointestinal nematodes. All trials included a group treated with derquantel-abamectin, and a negative control group. Nine trials included additional groups each treated with a single- or dual-active oral reference anthelmintic, selected from albendazole, levamisole, albendazole-levamisole, ivermectin, abamectin and moxidectin. A total of 838 animals were enrolled across all trials, and were randomly allocated to treatment groups within blocks defined by faecal nematode egg counts (FEC) pretreatment. On Day 0 derquantel-abamectin was administered orally at 1 ml/5 kg bodyweight (2 mg/kg derquantel, 0.2 mg/ kg abamectin), and each reference anthelmintic was given at the recommended label dose. Faecal samples were collected on Day 14 (+/- 1 day), to determine the percentage reduction in mean FEC for each anthelmintic tested. Larval differentiation was also performed post-treatment, to estimate efficacy at the genus level. Animals were weighed on or before Day 0, and on Day 14 (+/- 1 day) in 13 trials. RESULTS: The efficacy of derquantel-abamectin against mixed strongyle populations was > or =99.2%, based on the percentage reduction in geometric mean FEC. Nematodirus sp. was present in six trials at a level sufficient for efficacy calculations to be conducted; in all cases, the efficacy of derquantel-abamectin was 100%. In those trials where the efficacy of at least one reference anthelmintic was <95% against strongyles and/or Nematodirus sp., derquantel-abamectin was 100% effective. In five trials, the mean gain in bodyweight was significantly greater in the derquantel- abamectin group than the negative controls. CONCLUSIONS AND CLINICAL RELEVANCE: When administered orally at 1 ml/5 kg bodyweight, derquantel-abamectin is highly effective for the treatment of gastrointestinal nematodes in sheep, including populations of strongyles and Nematodirus sp. with resistance to one or more single- or dual-active anthelmintics. Derquantel-abamectin presents sheep producers with a unique opportunity to introduce a new class of anthelmintic to their nematode control programmes, with the added benefits offered by a combination anthelmintic.

Seroprevalence of Trypanosoma cruzi in rural Ecuador and clustering of seropositivity within households.

We performed a cross-sectional study of Trypanosoma cruzi seroprevalence in 14 communities in three provinces of Ecuador and estimated the magnitude of the association of seropositive individuals within households. A total of 3,286 subjects from 997 households were included. Seroprevalence was 5.7%, 1.0%, and 3.6% in subjects in the Manabí, Guayas, and Loja provinces, respectively. Seroprevalence increased with increasing age in Manabí and Guayas, whereas in Loja, the highest prevalence occurred in children

Household risk factors for Trypanosoma cruzi seropositivity in two geographic regions of Ecuador.

Few studies on the relationship between environmental factors and Trypanosoma cruzi transmission have been conducted in Ecuador. We conducted a cross-sectional study of household risk factors for T. cruzi seropositivity in 2 distinct geographical regions of Ecuador. Exposure information was collected via household surveys, and subjects were tested for serological evidence of T. cruzi infection. In total, 3,286 subjects from 997 households were included. In the coastal region, factors associated with seropositivity were living in a house with a palm roof (odds ratio [OR] = 2.63, 95% confidence interval, [1.61. 4.27]), wood walls (OR = 5.75 [2.04, 16.18]), or cane walls (OR = 2.81 11.31, 6.04]), and the presence of firewood in the peridomicile (OR = 2.48 [1.54, 4.01]). Accumulation of trash outside the home was associated with a reduced risk of seropositivity (OR = 0.25 [0.12, 0.51]). In the Andean region, living in a house with adobe walls was the only factor predictive of T. cruzi seropositivity. In conclusion, risk factors for T. cruzi transmission in Ecuador varied by geographic region, probably because of differing behavior of the triatomine vector species in each region. An understanding of the transmission dynamics of T. cruzi in a particular area is necessary for the development of effective Chagas disease control strategies in those areas.

Mathematical and statistical analysis of the Trypanosoma brucei slender to stumpy transition.

We propose a new model for the Stumpy Induction Factor-induced slender to stumpy transformation of Trypanosoma brucei gambiense cells in immunosuppressed mice. The model is a set of delay differential equations that describe the time-course of the infection. We fit the model, using a maximum-likelihood method, to previously published data on parasitaemia in four mice. The model is shown to be a good fit and parameter estimates and confidence intervals are derived. Our estimated parameter values are consistent with estimates from previous experimental studies. The model predicts the following. Slender cells can be classified as uncommitted, committed and dividing, and committed and non-dividing. A committed slender cell undergoes about 5 divisions before exiting the cell-cycle. Committed slender cells must produce SIF, and stumpy cells must not produce SIF. There are two mechanisms for differentiation, a background differentiation rate, and a SIF-concentration-dependent differentiation rate, which is proportional to SIF concentration. SIF has a half-life of about 1.4 h in mice. We also show, with suitable changes in the parameter values, that the model reflects behaviours seen in other host species and trypanosome strains.

Harmonisation of rat fetal skeletal terminology and classification. Report of the Third Workshop on the Terminology in Developmental Toxicology. Berlin, 14-16 September 2000.

The initial efforts of the Federal Institute for Health Protection of Consumers and Veterinary Medicine (BgVV) and the Free University of Berlin to standardise terminology in the field of developmental toxicology began in 1995. Procedures were undertaken to harmonise the terminology used by the International Federation of Teratology Societies (IFTS) and the International Programme on Chemical Safety (IPCS). This article reflects these activities and is a report on the Third Workshop on the Terminology in Developmental Toxicology held in September 2000. This Workshop served as a forum to discuss the results of a survey on the classification of skeletal anomalies that had been previously sent to scientists active in the field. Although high agreement was reached among the evaluators for several terms, the use of a number of terms was rather variable. Therefore, the discussions at the workshop among the experts from research institutions, regulatory agencies, and industry were mainly focussed on those terms for which there was disagreement and/or uncertainties and the possible reasons. Pictures provided by the participants for the illustration of "grey zone" anomalies constituted the basis for detailed discussions. In many of the cases with lower agreement, decisions were facilitated by the provision of the corresponding picture. The main reasons for lower agreement were imprecise terms, insufficient knowledge on postnatal consequences, theoretical terms that are unlikely to occur in isolation, and the possibility of observing a range of severity that might be decisive for the classification of either a malformation or variation. The attendees concluded that "grey-zone" anomalies will never disappear completely and that for the assessment, the grade of severity and/or the frequency of the observation can be decisive for the terminology chosen. A Joint IPCS/IFTS Project was proposed to further consensus of terminology and classification and to link these anomalies to pictures at different skeletal sites. In order to support the harmonisation of regulatory decisions, it was proposed to establish a "Clearinghouse" System under the umbrella of the IPCS. The Clearinghouse could be contacted either by the regulatory authorities or by any company to clarify their queries, particularly with regard to registration or authorisation processes. Finally, it was recommended to also carry out a similar survey on "soft tissue anomalies" and "external findings." The results of this survey will be discussed at a Joint IPCS/IFTS Workshop in Berlin in 2002.

African trypanosomiasis research: 100 years of progress, but questions and problems still remain.

Past and present progress in our understanding of African trypanosomiasis is briefly reviewed. Although tremendous scientific strides have been achieved, an epidemic of the disease is currently underway. Three areas of research which are believed necessary for the control of African trypanosomiasis are discussed. It is suggested that a better understanding of the host-parasite relationship is essential; more emphasis and a broader approach to drug development is required; and finally, further research into the socio-economic aspects of African trypanosomiasis is urgently needed before the human disease can again be controlled.

Harmonization of cancer and noncancer risk assessment: proceedings of a consensus-building workshop.

Significant advancements have been made toward the use of all relevant scientific information in health risk assessments. This principle has been set forth in risk-assessment guidance documents of international agencies including those of the World Health Organization's International Programme on Chemical Safety, the U.S. Environmental Protection Agency, and Health Canada. Improving the scientific basis of risk assessment is a leading strategic goal of the Society of Toxicology. In recent years, there has been a plethora of mechanistic research on modes of chemical toxicity that establishes mechanistic links between noncancer responses to toxic agents and subsequent overt manifestations of toxicity such as cancer. The research suggests that differences in approaches to assessing risk of cancer and noncancer toxicity need to be resolved and a common broad paradigm for dose-response assessments developed for all toxicity endpoints. In November 1999, a workshop entitled "Harmonization of Cancer and Noncancer Risk Assessment" was held to discuss the most critical issues involved in developing a more consistent and unified approach to risk assessment for all endpoints. Invited participants from government, industry, and academia discussed focus questions in the areas of mode of action as the basis for harmonization, common levels of adverse effect across toxicities for use in dose-response assessments, and scaling and uncertainty factors. This report summarizes the results of those discussions. There was broad agreement, albeit not unanimous, that current science supports the development of a harmonized set of principles that guide risk assessments for all toxic endpoints. There was an acceptance among the participants that understanding the mode of action of a chemical is ultimately critical for nondefault risk assessments, that common modes of action for different toxicities can be defined, and that our approach to assessing toxicity should be biologically consistent.

Innate and acquired resistance to African trypanosomiasis.

The review discusses the current field status of human and bovine trypanosomiases, and focuses on the molecular basis of innate and acquired control of African trypanosomes in people, cattle, and Cape buffalo.

Workshop to identify critical windows of exposure for children's health: reproductive health in children and adolescents work group summary.

This work group report addresses the central question: What are the critical windows during development (preconception through puberty) when exposure to xenobiotics may have the greatest adverse impact on subsequent reproductive health? The reproductive system develops in stages, with sex-specific organogenesis occurring prenatally and further maturational events occurring in the perinatal period and at puberty. Complex endocrine signals as well as other regulatory factors (genetics, growth factors) are involved at all stages. Evidence from animal models and human studies indicates that many specific events can be perturbed by a variety of toxicants, with endocrine-mediated mechanisms being the more widely studied. Prioritized research needs include basic studies on the cellular-molecular and endocrine regulation of sexual differentiation and development; increased efforts regarding potential adverse effects on development in females, including breast development; expanded animal studies on different classes of chemicals, comparing responses during development (prenatal and postnatal) with responses in adults; and, more extensive explorations regarding the reproductive biology and toxicology of puberty in humans.

Basal activity of the hypothalamic-pituitary-adrenal axis and cognitive function in anorexia nervosa.

Anorexia nervosa is associated with abnormalities in neuroendocrine function including sustained hypercortisolism, which has been shown elsewhere to be associated with impairment of function in learning, memory and attention. Cognitive impairment has also been observed in anorexia nervosa. These effects may be mediated in part through cortisol effects on the hippocampus, which is dense with glucocorticoid receptors. We investigated the association between cortisol levels and cognitive function in anorexia nervosa by measuring both 24-hour urinary cortisol counts and performance on tasks of learning, memory and attention in patients suffering from the disorder. Cortisol secretion was shown to be significantly higher in the patient group than in a matched control group and patients were also shown to be impaired in memory and attention. However, no correlations were found between the cognitive deficits and cortisol measures. It is suggested that more sensitive profiling of cortisol levels throughout the circadian cycle may be useful in future studies of cognitive function in anorexia nervosa.

A revised arithmetic model of long slender to short stumpy transformation in the African trypanosomes.

An arithmetic model that closely approximates an African trypanosome infection in immunosuppressed mice is presented. The final model was based on an examination of the following parameters: the rate of long slender to short stumpy transition, the maximum percentage of long slender to short stumpy stages that can be induced, the survival time or half life of the short stumpy stage in vivo, and the rate (%) of long slender to short stumpy stage transition following the peak in transformation. The model is based on the assumption that the long slender to short stumpy transition is parasite population dependent and that in mice the long slender to short stumpy transition only begins when the trypanosome population reaches a density of 1 x 10(7) trypanosomes/ml. The model predicts that the parasitemia during the first several days of an infection is controlled solely by the kinetics of the transition of the dividing long slender stage to the nondividing short stumpy stage. It was not necessary to include in the model the host's immune response in order to simulate the early growth kinetics of pleomorphic trypanosomes in infected mice.

Anti-Trypanosoma brucei activity of nonprimate zoo sera.

Constitutive anti-Trypanosoma brucei subsp. brucei S 427 clone 1 and 22 activities were evaluated in sera from 22 species of nonprimate mammals. The sera fell into 5 categories. Sera from Cape buffalo, giraffe, and greater kudu showed a concentration-dependent inhibition of replication of the 2 clones of organisms, which was dependent on the presence of xanthine oxidase. Sera from warthog and springbok also severely limited trypanosome replication but lacked xanthine oxidase. Their antitrypanosome activity was inactivated by heating at 56 C for 30 min but not affected by absorbing with trypanosomes at 4 C. Sera from lion and leopard showed a concentration-dependent inhibition of the growth of T. brucei S427 clone 1 organisms, but not clone 22 organisms. These sera lacked xanthine oxidase. Their anti-T. brucei S 427 clone 1 activity was inactivated by heating at 56 C for 30 min but not removed by absorbing with trypanosomes. Serum from Grant's gazelle prevented replication of both T. brucei clones, lacked xanthine oxidase, and was not affected by heating at 56 C. Sera from waterbuck, Thompson's gazelle, sitatunga, Cape hartebeeste, gerenuk, Grant's zebra, cow, several cat, cougar, bobcat, and domestic cat were fully supportive of trypanosome replication irrespective of concentration tested up to a maximum of 48% v/v in culture medium. Sera from different individuals of the same mammal species had similar effects on trypanosomes, and samples collected from the same individual at different times also had similar activities indicating species-specific stable expression, or lack thereof, of constitutive serum antitrypanosome components.

Increased excretion of aromatic amino acid catabolites in animals infected with Trypanosoma brucei evansi.

Aromatic amino acid catabolism by Trypanosoma brucei evansi was investigated in vivo using C3HeB/FeJ mice. The major catabolites detected by gas chromatography in the urines of infected animals were phenylpyruvic acid, 4-hydroxyphenylpyruvic acid, and indole-3-pyruvic acid. Identity of each compound was confirmed by gas chromatography-mass spectrometry. Concentrations of catabolites in urine of infected mice were correlated with parasitemia and returned to normal following suramin treatment. Other aromatic amino acid metabolites, including indole-3-acetic acid, indole-3-lactic acid, and 4-hydroxyphenyllactic acid, were detected in urine from infected animals by gas chromatography mass spectrometry, although quantities were too low to be quantified reproducibly. Both phenylpyruvic acid and 4-hydroxyphenylpyruvic acid were also detected in urine of dogs and donkeys experimentally infected in Egypt with a recent field isolate of T. b. evansi. Tryptophan metabolites could not be assayed in dog and urine samples because formalin, which degraded the indole acids, had to be added before the samples could be imported into the U.S. Finally, concentrations of urinary catabolites during infection were correlated with the tyrosine aminotransferase activity in infected mouse sera.

Ascaris lumbricoides intensity in relation to environmental, socioeconomic, and behavioral determinants of exposure to infection in children from southeast Madagascar.

Ascaris lumbricoides worm counts were examined as the outcome products of exposure proxy variables. A survey of 663 children, 4-10 yr old, living in southeastern Madagascar revealed prevalences of 93% for A. lumbricoides, 55% for Trichuris trichiura, and 27% for hookworm. Worm expulsions were conducted on 428 of these children; the data revealed an overdispersed distribution of A. lumbricoides, with an arithmetic mean of 19.2 worms per child. A concurrent socioeconomic household survey was conducted by visitation and interview. Exposure to infection was assessed by environmental, demographic, behavioral, and socioeconomic indicators. Ascaris lumbricoides aggregations were associated with gender, housing style, ethnicity, and agricultural factors. The results suggest that exposure and infection are ubiquitous in this child population, and that A. lumbricoides intensity is influenced by gender-related behavioral and environmental factors that contribute to exposure.

Diagnosis of trypanosomiasis in experimental mice and field-infected camels by detection of antibody to trypanosome tyrosine aminotransferase.

Sera from animals with acute and chronic Trypansoma evansi infections were examined directly for trypanosome tyrosine aminotransferase activity and indirectly for their ability to inhibit tyrosine aminotransferase activity. It was shown that sera from acutely infected mice and camels with high parasitemias contained significant levels of trypanosome tyrosine aminotransferase activity. In contrast, the sera from chronically infected mice and camels did not contain significant tyrosine aminotransferase activity, but they were able to neutralize the enzyme activity in trypanosome homogenates. The sera from camels with other pathological conditions did not neutralize this enzyme activity. It is suggested that the inhibitory factor in the chronic sera is antibody. The potential use of the direct enzyme assay and the indirect neutralization assay as diagnostic tools are discussed. Finally, the use of these assays to distinguish between early (acute) and late (chronic) infections are also suggested.