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Symposium overview: mechanism of action of nicotine on neuronal acetylcholine receptors, from molecule to behavior.

Nicotine has long been known to interact with nicotinic acetylcholine (ACh) receptors since Langley used it extensively to chart sympathetic ganglia a century ago. It has also been used as an effective insecticide. However, it was not until the 1990s that the significance of nicotine was increasingly recognized from the toxicological, pharmacological, and environmental points of view. This is partly because studies of neuronal nicotinic ACh receptors are rapidly emerging from orphan status, fueled by several lines of research. Since Alzheimer's disease is known to be associated with down-regulation of cholinergic activity in the brain, a variety of nicotine derivatives are being tested and developed for treatment of the disease. Public awareness of the adverse effects of nicotine has reached the highest level recently. Since insect resistance to insecticides is one of the most serious issues in the pest-control arena, it is an urgent requirement to develop new insecticides that act on target sites not shared by the existing insecticides. The neuronal nicotinic ACh receptor is one of them, and new nicotinoids are being developed. Thus, the time is ripe to discuss the mechanism of action of nicotine from a variety of angles, including the molecular, physiological, and behavioral points of view. This Symposium covered a wide area of nicotine studies: genetic, genomic, and functional aspects of nicotinic ACh receptors were studied, as related to anthelmintics and insecticides; interactions between ethanol and nicotine out the ACh receptor were analyzed, in an attempt to explain the well-known heavy drinker-heavy smoker correlation; the mechanisms that underlie the desensitization of ACh receptors were studied as related to nicotine action; selective pharmacological profiles of nicotine, and descriptions of some derivatives were described; and chronic nicotine infusion effects on memory were examined using animal models.

Animals↗

The common insecticides cyfluthrin and chlorpyrifos alter the expression of a subset of genes with diverse functions in primary human astrocytes.

Given the widespread use of insecticides in the environment, it is important to perform studies evaluating their potential effects on humans. Organophosphate insecticides, such as chlorpyrifos, are being phased out; however, the use of pyrethroids in household pest control is increasing. While chlorpyrifos is relatively well studied, much less is known about the potential neurotoxicity of cyfluthrin and other pyrethroids. To gain insights into the neurotoxicity of cyfluthrin, we compared and evaluated the toxicity profiles of chlorpyrifos and cyfluthrin in primary human fetal astrocytes. We found that at the same concentrations, cyfluthrin exerts as great as, or greater toxic effects on the growth, survival, and proper functioning of human astrocytes. By using microarray gene expression profiling, we systematically identified and compared the potential molecular targets of chlorpyrifos and cyfluthrin, at a genome-wide scale. We found that chlorpyrifos and cyfluthrin affect a similar number of transcripts. These targets include molecular chaperones, signal transducers, transcriptional regulators, transporters, and those involved in behavior and development. Further computational and biochemical analyses show that cyfluthrin and chlorpyrifos upregulate certain targets of the interferon-gamma and insulin-signaling pathways and that they increase the protein levels of activated extracellular signal-regulated kinase 1/2, a key component of insulin signaling; interleukin 6, a key inflammatory mediator; and glial fibrillary acidic protein, a marker of inflammatory astrocyte activation. These results suggest that inflammatory activation of astrocytes might be an important mechanism underlying neurotoxicity of both chlorpyrifos and cyfluthrin.

Apoptosis↗

Mortality of pesticide applicators.

Information has been obtained on a cohort of professional pesticide applicators which will be followed prospectively. The cohort consists of 16,126 males employed for three months or more between 1967 and 1976 by any of three nationwide pest control companies. Deaths which occurred between 1967 and 1976 were reported by the Social Security Administration as a result of a search of its records. Overall, 311 deaths were ascertained, giving a standardized mortality ration (SMR) of 84. SMRs were over 100 for three causes of death--cancer of the lung (115), cancer of the skin (173) and cancer of the bladder (277). The confidence intervals of two of these ratios include 100, and the observed numbers therefore do not differ significantly from those expected. For bladder cancer the excess is on the border of statistical significance (p less than 0.05). The excess of deaths from lung cancer was not seen for applicators classified as termite control operators, a group more likely to be exposed to chlordane and heptachlor. There were significantly low SMRs for cancer of the digestive organs (46) and for other diseases of the digestive (55) and respiratory (29) systems. Deaths from cerebrovascular disease were also less than expected, though not significantly so.

Cerebrovascular Disorders↗

Space, time and persistence of virulent pathogens.

Epidemiological theory predicts that pathogens of high virulence should not become endemic. We show, using an empirically based lattice map model, that a pathogen that is too virulent to persist if its host population is spatially well mixed, can persist if the host population is spatially distributed, because of internally generated complex spatial dynamics, provided that the area occupied by the host population is sufficiently large. The dynamics are not an artefact of spatial or temporal discretization. The results uncover a mechanism for the persistence of virulent pathogens, suggesting a means by which pathogens of high virulence could achieve sustained as well as short-term biological pest control.

Animals↗

Overcoming cytoplasmic incompatibility in Drosophila.

The endocellular microbe Wolbachia pipientis infects a wide variety of invertebrate species, in which its presence is closely linked to a form of reproductive failure termed cytoplasmic incompatibility (CI). CI renders infected males unable to father offspring when mated to uninfected females. Because CI can dramatically affect fitness in natural populations, mechanisms that abate CI can have equally large impacts on fitness. We have discovered that repeated copulation by Wolbachia-infected male Drosophila simulans significantly diminishes CI. Repeated copulation does not prevent Wolbachia from populating developing spermatids, but may reduce the time during spermatogenesis when Wolbachia can express CI. This restoration of fertility in premated infected males could have important implications for Wolbachia transmission and persistence in nature and for its exploitation as an agent of biological pest control.

Animals↗

Novel genetic basis of field-evolved resistance to Bt toxins in Plutella xylostella.

Insecticidal toxins from Bacillus thuringiensis (Bt) are widely used to control pest insects, but evolution of resistance threatens their continued efficacy. The most common type of Bt resistance ('Mode 1') is characterized by recessive inheritance, > 500-fold resistance to at least one Cry1A toxin, negligible cross-resistance to Cry1C, and reduced binding of Bt toxins to midgut membrane target sites. Mutations affecting a Cry1A-binding midgut cadherin protein are linked to laboratory-selected Mode 1 resistance in Heliothis virescens and Pectinophora gossypiella. Here we show that field-evolved Mode 1 resistance in the diamondback moth, Plutella xylostella, has a different genetic basis, indicating that screening for resistance in the field should not be restricted to a previously proposed DNA-based search for cadherin mutations.

Animals↗

Field evaluation of four biorational larvicide formulations against Anopheles albimanus in Honduras.

Four biorational larvicide formulations: Teknar (B.t.i.), Arosurf MSF (Monomolecular Surface Film), Arosurf MSF combined with Teknar, and SAN-809-I ([s]-methoprene combined with B.t.i.), were evaluated against naturally occurring populations of the malaria vector mosquito Anopheles albimanus Wiedemann in Honduras. All formulations reduced the mean number of larvae per sample area to 0 within 48 h after treatment, and gave significant (P less than 0.05) control when compared with similar untreated areas for at least 10 days after treatment. It is concluded that each of these four formulations can be used effectively to control the larvae of An. albimanus in Honduras.

Animals↗

Optimal life stage for radiation sterilization of Anopheles males and their fitness for release.

Mating by male Anopheles mosquitoes (Diptera: Culicidae) was evaluated in the laboratory to assess fitness effects of radio-sterilization applied during different life stages of the malaria vectors An. stephensi Liston and An. gambiae Giles sensu stricto. After reproductive sterilization by gamma-irradiation (120 Gy), equal proportions of sterile and fertile (unirradiated) male adults were released into cages with virgin females and allowed to compete for matings. Radio-sterilization was applied when the males were pupae aged 0-7 h or 24-32 h, or adults aged <24 h or 24-55 h. After being radio-sterilized in the adult stage, males of both species competed effectively with unirradiated males, whereas those sterilized in the pupal stage obtained significantly fewer matings than unirradiated males from the same cohort. There was no evidence of females obtaining multiple inseminations. These findings emphasize the need to radio-sterilize males as adults in order to minimize the fitness cost. Such males may be intended for sterile insect technique population suppression or for trial releases of transgenic anophelines.

Animals↗

Infiltration by alien predators into invertebrate food webs in Hawaii: a molecular approach.

Abstract Alien invertebrate predators have been introduced to Hawaii to control pests, particularly in lowland areas where most crops are grown. We developed techniques for assessing the impact of these predators on native food webs in relatively pristine upland areas where, it was hypothesized, few lowland predators might be found. Predator densities were assessed along transects within the Alakaii Swamp on Kaua'i. The most numerous alien biocontrol agents found were Halmus chalybeus (Coccinellidae), a species known to feed on Lepidoptera eggs. Laboratory experiments were conducted using two genera of endemic Lepidoptera, Scotorythra and Eupithecia (Geometridae), that are of considerable conservation value, the former because of its recent speciation across Hawaii, the latter for its unique predatory larvae. Techniques were developed for detecting Lepidoptera DNA within the guts of alien predators using prey-specific PCR primers. General primers amplified fragments of the mitochondrial cytochrome oxidase I gene from beetles and Lepidoptera. The sequences were aligned and used successfully to design target-specific primers for general detection of the remains of Geometridae and for particular species, including Scotorythra rara and Eupithecia monticolans. DNA fragments amplified were short [140-170 base pairs (bp)], optimizing detection periods following prey ingestion. Trials using the introduced biocontrol agent Curinus coeruleus (Coccinellidae) demonstrated detection of Lepidoptera DNA fragments = 151 bp in 85-100% of beetles after 24 h digestion of an early instar larva. This study provides a framework for future use of molecular gut analysis in arthropod conservation ecology and food web research with considerable potential for quantifying threats to endemic species in Hawaii and elsewhere.

Amino Acid Sequence↗

Uptake of Bt endotoxins by nontarget herbivores and higher order arthropod predators: molecular evidence from a transgenic corn agroecosystem.

The planting of transgenic crops expressing Bacillus thuringiensis endotoxins is widespread throughout the world; the prolific increase in their application exposes nontarget organisms to toxins designed to control pests. To date, studies have focused upon the effects of Bt endotoxins on specific herbivores and detritivores, without consideration of their persistence within arthropod food webs. Here, we report the first quantitative field evaluation of levels of Bt endotoxin within nontarget herbivores and the uptake by higher order arthropods. Antibody-based assays indicated significant quantities of detectable Cry1Ab endotoxin within nontarget herbivores which feed on transgenic corn (including the corn flea beetle, Chaetocnema pulicaria, Japanese beetle, Popillia japonica and southern corn rootworm, Diabrotica undecimpunctata howardi). Furthermore, arthropod predators (Coccinellidae, Araneae, and Nabidae) collected from these agroecosystems also contained significant quantities of Cry1Ab endotoxin indicating its movement into higher trophic levels. This uptake by predators is likely to have occurred by direct feeding on plant material (in predators which are facultatively phytophagous) or the consumption of arthropod prey which contained these proteins. These data indicate that long-term exposure to insecticidal toxins occurs in the field. These levels of exposure should therefore be considered during future risk assessments of transgenic crops to nontarget herbivores and arthropod predators.

Agriculture↗

Communities of arbuscular mycorrhizal fungi in arable soils are not necessarily low in diversity.

Communities of arbuscular mycorrhizal fungi (AMF) in five agricultural field sites of different management intensities were studied. Variable regions of the ribosomal RNA genes were used to detect and identify AMF directly within colonized roots. Roots from a continuous maize monoculture showed low AMF diversity, in agreement with previous reports on molecular diversity of AMF in agricultural soils. In contrast, a substantially higher diversity of AMF was found throughout the long term 'DOK' field experiment, where organic and conventional agricultural practices have been compared side by side since 1978. In this experiment, a 7-year crop rotation is performed under lower levels of inorganic fertilizer input and chemical pest control. These results are in good agreement with analyses of the spore community previously conducted in these field sites. In a third site, an organically managed leek field with soil of very high phosphate content reflecting the highly intensive conventional field history and intensive tillage, we detected a low-diversity community comparable to the maize monoculture. In addition to fungi from Glomus group A, which have previously been reported to dominate arable soils, we regularly found members of the genera Scutellospora, Paraglomus and Acaulospora. The genus Acaulospora was shown to occur more frequently early in the growing season, suggesting that the life history strategy of AMF may influence the active community at a given time. These data show that the diversity of AMF is not always low in arable soils. Furthermore, low-input agriculture involving crop rotation may provide better conditions to preserve AMF diversity, by preventing the selection for the few AMF taxa tolerating high nutrient levels.

Agriculture↗

Suppression of exposure to malaria vectors by an order of magnitude using microbial larvicides in rural Kenya.

OBJECTIVE: To determine the contribution larviciding could make to reduce the burden of malaria, by conducting a trial of microbial larvicides in a 4.5 km2 area in and around a large village in rural western Kenya. METHOD: The abundance of immature and adult mosquitoes was monitored for 12 months under baseline conditions. Then microbial larval control was implemented for 28 months. After the intervention, the abundance of immature and adult mosquitoes was monitored for a further 12 months. RESULTS: Of the 419 mosquito larval habitats identified, 336 (80%) originated from human activities. Application of Bacillus thuringiensis var. israelensis and Bacillus sphaericus larvicides reduced the proportion of aquatic habitats containing Anopheles larvae from 51% during non-intervention periods to 7% during the intervention. The occurrence of late instar Anopheles in habitats was reduced from 39% and 33% in pre-intervention and post-intervention periods to 0.6% during intervention. Overall, larviciding reduced Anopheles larval density by 95% and human exposure to bites from adults by 92%. The estimated cost of providing this protection to the human population in the study area was less than US$ 0.90/person/year. CONCLUSION: Appropriately applied microbial larvicides can substantially and cost-effectively reduce human exposure to malaria in rural sub-Saharan Africa.

Animals↗

Neutral N-glycans of the gastropod Arion lusitanicus.

The neutral N-glycan structures of Arion lusitanicus (gastropod) skin, viscera and egg glycoproteins were examined after proteolytic digestion, release of the glycans from the peptides, fluorescent labelling with 2-aminopyridine and fractionation by charge, size and hydrophobicity to obtain pure glycan structures. The positions and linkages of the sugars in the glycan were analysed by two dimensional HPLC (size and hydrophobicity) and MALDI-TOF mass spectrometry before and after digestion with specific exoglycosidases. The most striking feature in the adult tissues was the high amount of oligomannosidic and small paucimannosidic glycans terminated with 3-O-methylated mannoses. The truncated structures often contained modifications of the inner core by beta1,2-linked xylose to the beta-mannose residue and/or an alpha-fucosylation (mainly alpha1,6-) of the innermost GlcNAc residue. Skin and viscera showed predominantly the same glycans, however, in different amounts. Traces of large structures carrying 3-O-methylated galactoses were also detected. The egg glycans contained mainly (approximately 75%) oligomannosidic structures and some paucimannosidic structures modified by xylose or alpha1,6-fucose, but in this case no methylation of any monosaccharide was detected. Thus, gastropods seem to be capable of producing many types of structures ranging from those typical in human to structures similar to those found in nematodes, and therefore will be a valuable model to understand the regulation of glycosylation. Furthermore, this opens the way for using this organism as a host for the production of recombinant proteins. The detailed knowledge on glycosylation also may help to identify targets for pest control.

Aminopyridines↗

Variation in cutaneous sensation between synthetic pyrethroid insecticides.

Synthetic pyrethroids are potent insecticides, utilized for food protection and general pest control. Numerous investigations have indicated that ultra-low volume applications are effective and may eliminate the developing problem of resistance to the currently utilized insecticides. The most prominent health symptom that accompanies topical contact with these agents appears to be a cutaneous sensation, paresthesia. In this investigation, a substantial difference in the degree of paresthesia was noted between the formulated grade of 4 synthetic pyrethroids. Also, dl-alpha tocopheryl acetate was statistically validated as an efficacious therapeutic agent for cutaneous exposure to these insecticides.

Administration, Topical↗