Toxicity Testing in the 21st Century: Defining New Risk Assessment Approaches Based on Perturbation of Intracellular Toxicity Pathways

The approaches to quantitatively assessing the health risks of chemical exposure have not changed appreciably in the past 50 to 80 years, the focus remaining on high-dose studies that measure adverse outcomes in homogeneous animal populations. This expensive, low-throughput approach relies on conservative extrapolations to relate animal studies to much lower-dose human exposures and is of questionable relevance to predicting risks to humans at their typical low exposures. It makes little use of a mechanistic understanding of the mode of action by which chemicals perturb biological processes in human cells and tissues. An alternative vision, proposed by the U.S. National Research Council (NRC) report Toxicity Testing in the 21^st Century: A Vision and a Strategy, called for moving away from traditional high-dose animal studies to an approach based on perturbation of cellular responses using well-designed in vitro assays. Central to this vision are (a) “toxicity pathways” (the innate cellular pathways that may be perturbed by chemicals) and (b) the determination of chemical concentration ranges where those perturbations are likely to be excessive, thereby leading to adverse health effects if present for a prolonged duration in an intact organism. In this paper we briefly review the original NRC report and responses to that report over the past 3 years, and discuss how the change in testing might be achieved in the U.S. and in the European Union (EU). EU initiatives in developing alternatives to animal testing of cosmetic ingredients have run very much in parallel with the NRC report. Moving from current practice to the NRC vision would require using prototype toxicity pathways to develop case studies showing the new vision in action. In this vein, we also discuss how the proposed strategy for toxicity testing might be applied to the toxicity pathways associated with DNA damage and repair.     

Structural insight into the differential effects of omega-3 and omega-6 fatty acids on the production of Abeta peptides and amyloid plaques

Several studies have shown the protective effects of dietary enrichment of various lipids in several late-onset animal models of Alzheimer Disease (AD); however, none of the studies has determined which structure within a lipid determines its detrimental or beneficial effects on AD. High-sensitivity enzyme-linked immunosorbent assay (ELISA) shows that saturated fatty acids (SFAs), upstream omega-3 FAs, and arachidonic acid (AA) resulted in significantly higher secretion of both Aβ 40 and 42 peptides compared with long chain downstream omega-3 and monounsaturated FAs (MUFA). Their distinct detrimental action is believed to be due to a structural template found in their fatty acyl chains that lack SFAs, upstream omega-3 FAs, and AA. Immunoblotting experiments and use of APP-C99-transfected COS-7 cells suggest that FA-driven altered production of Aβ is mediated through γ-secretase cleavage of APP. An early-onset AD transgenic mouse model expressing the double-mutant form of human amyloid precursor protein (APP); Swedish (K670N/M671L) and Indiana (V717F), corroborated in vitro findings by showing lower levels of Aβ and amyloid plaques in the brain, when they were fed a low fat diet enriched in DHA. Our work contributes to the clarification of aspects of structure-activity relationships.     

Opiine parasitoids (Hymenoptera: Braconidae) of tropical fruit flies (Diptera: Tephritidae) of the Australian and South Pacific region

Opiine wasps are parasitoids of dacine fruit flies, the primary horticultural pests of Australia and the South Pacific. A taxonomic synopsis and distribution and host records (44% of which are new) for each of the 15 species of dacine-parasitizing opiine braconids found in the South Pacific is presented. Species dealt with are Diachasmimorpha hageni (Fullaway), D. kraussii (Fullaway), D. longicaudata (Ashmead), D. tryoni (Cameron), Fopius arisanus (Sonan), F. deeralensis (Fullaway), F. ferrari Carmichael & Wharton sp. n., F. illusorius (Fischer) comb. n., F. schlingeri Wharton, Opius froggatti Fullaway, Psyttalia fijiensis (Fullaway), P. muesebecki (Fischer), P. novaguineensis (Szépligeti) and Utetes perkinsi (Fullaway). A potentially undescribed species, which may be a colour morph of F. vandenboschi (Fullaway), is diagnosed but not formally described. Fopius vandenboschi sensu stricto, Diachasmimorpha fullawayi Silvestri, Psyttalia concolor Szépligeti and P. incisi Silvestri have been liberated into the region but are not considered to have established: a brief diagnosis of each is included. Biosteres illusorius Fischer is formally transferred to the genus Fopius. A single opiine specimen reared from a species of Bactrocera (Bulladacus) appears to be Utetes albimanus (Szépligeti), but damage to this specimen and to the holotype (the only previously known specimen) means that this species remains unconfirmed as a fruit fly parasite: a diagnosis of U. cf. albimanus is provided. Psyttalia novaguineensis could not be adequately separated from P. fijiensis using previously published characterizations and further work to resolve this complex is recommended. A key is provided to all taxa.     

Evidence for paralytic shellfish poisons in the freshwater cyanobacterium Lyngbya wollei (Farlow ex Gomont) comb. nov.

Lyngbya wollei (Farlow ex Gomont) comb. nov., a perennial mat-forming filamentous cyanobacterium prevalent in lakes and reservoirs of the southeastern United States, was found to produce a potent, acutely lethal neurotoxin when tested in the mouse bioassay. Signs of poisoning were similar to those of paralytic shellfish poisoning. As part of the Tennessee Valley Authority master plan for Guntersville Reservoir, the mat-forming filamentous cyanobacterium L. wollei, a species that had recently invaded from other areas of the southern United States, was studied to determine if it could produce any of the known cyanotoxins. Of the 91 field samples collected at 10 locations at Guntersville Reservoir, Ala., on the Tennessee River, over a 3-year period, 72.5% were toxic. The minimum 100% lethal doses of the toxic samples ranged from 150 to 1,500 mg kg of lyophilized L. wollei cells-1, with the majority of samples being toxic at 500 mg kg-1. Samples bioassayed for paralytic shellfish toxins by the Association of Official Analytical Chemists method exhibited saxitoxin equivalents ranging from 0 to 58 micrograms g (dry weight)-1. Characteristics of the neurotoxic compound(s), such as the lack of adsorption by C18 solid-phase extraction columns, the short retention times on C18 high-performance liquid chromatography (HPLC) columns, the interaction of the neurotoxins with saxiphilin (a soluble saxitoxin-binding protein), and external blockage of voltage-sensitive sodium channels, led to our discovery that this neurotoxin(s) is related to the saxitoxins, the compounds responsible for paralytic shellfish poisonings. The major saxitoxin compounds thus far identified by comparison of HPLC fluorescence retention times are decarbamoyl gonyautoxins 2 and 3. There was no evidence of paralytic shellfish poison C toxins being produced by L. wollei. Fifty field samples were placed in unialgal culture and grown under defined culture conditions. Toxicity and signs of poisoning for these laboratory-grown strains of L. wollei were similar to those of the field collection samples.     

Circannual rhythms of ground squirrels: a test of the frequency demultiplication hypothesis

The frequency demultiplication hypothesis (FDH) posits that circannual rhythms are generated from circadian cycles by frequency transformation to the lower-frequency rhythm. To test the FDH, we determined the periods of the circannual body mass and estrous cycles of golden-mantled ground squirrels with circadian locomotor activity rhythms entrained to 23-, 24-, or 25-hr days (T-cycles). Circannual period length did not differ among squirrels entrained to the different T-cycles; intergroup ranges were 298-314 days and 303-312 days, respectively, for body mass and estrus. These results are not consistent with the FDH and suggest instead that separate mechanisms generate circadian and circannual rhythms. In ground squirrels the circannual system influences circadian organization, but a reciprocal influence of circadian on circannual rhythms has yet to be demonstrated.     

Fungi from Alberta rodents

Summary The occurrence, by years, ofE. parva in Alberta rodents is presented in tabular form covering the period 1946 to 1959. The infection rate was 5.1.% in 1946, 3.3% in 1957 and less than 1% thereafter.It was found that the lungs of Alberta rodents have a characteristic mold flora comprised chiefly of:Penicillium spp., Beauveria bassiana, Aspergillus spp. and perhapsScopulariopsis brevicaule, Cladosporium spp. andChrysosporium pannorum. Trichophyton mentagrophytes and anotherTrichophyton were isolated from the livers, spleens or lungs of six rodents. Phycomyces blakesleeanus was isolated from the lungs of two rodents trapped in northern Alberta.     

Microsporum nanum infection in Alberta

Summary Microsporum nanum Fuentes was isolated from a ringworm lesion on the back of the hand of a seven year old Alberta schoolgirl. This is the first record ofM. nanum in Canada and the first infection reported from the northern part of the temperate zone.