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The house dust mite allergen Der p1 catalytically inactivates alpha 1-antitrypsin by specific reactive centre loop cleavage: a mechanism that promotes airway inflammation and asthma.

Der p1, a cysteine proteinase derived from the house dust mite (HDM) Dermatophagoides pteronyssinus, is a major component of the allergic immune response in HDM atopic individuals. Recent evidence suggests that cysteine proteinase activity is important in the disease process as it increases the permeability of the allergen in the respiratory tract and disrupts the regulation of IgE synthesis. Der p1 is found in high concentrations in the faecal pellets of mites which are aerosolised and inhaled via the respiratory tract. The serine proteinase inhibitor, alpha 1-antitrypsin, protects the lower respiratory tract against damage by proteinases released in the lung during inflammation. Der p1 catalytically inactivates alpha 1-antitrypsin by a thiol-dependent mechanism involving specific cleavage of the reactive centre loop and we propose that this mechanism may be important in the pathogenesis of asthma.

Allergens↗

Acid-induced polymerization of the group 5 mite allergen from Dermatophagoides pteronyssinus.

House dust mites are the most important source of indoor allergens and cause allergic diseases. Our studies here suggest that the group 5 allergen from Dermatophagoides pteronyssinus (Der p 5) is monomeric at neutral pH, but forms filaments at low pH. Circular dichroism measurements show Der p 5 is a helical protein, and the protein sequence reveals Der p 5 contains coiled-coil helices. The acid-induced filament assembly could be explained in part by the high content of charged residues (40%) in the coiled-coil structure. Interestingly, some of the known Dermatophagoides allergens also contain a heptad repeat, which could potentially form coiled coils. Therefore, coiled-coil helices may be one of the common structural motifs of mite allergens that contribute to their allergenicity.

Allergens↗

Phylogeny of feather mite subfamily Avenzoariinae (Acari: Analgoidea: Avenzoariidae) inferred from combined analyses of molecular and morphological data.

Phylogenetic relationships among feather mites of the subfamily Avenzoariinae (Acari: Analgoidea: Avenzoariidae) were reconstructed by parsimony analysis of a combined data matrix. We analyzed 41 morphological characters and 246 molecular characters from a fragment of the 16S rDNA. Morphological trees were well supported at deep branches (genera and above), but showed much less support and resolution within genera. Molecular analyses produced trees with better resolution and support on terminal branches and worse support on basal branches. I(MF) index for the combined matrix pointed to the significant congruence of both data subsets with the whole of the data. The topology of the combined tree was close to the morphological tree in the deep branches and had well-resolved terminal branches as in the molecular tree. This suggests a considerable level of complimentarity between the two data sets. An analysis of association patterns of the mites and their hosts was conducted based on the results of the combined analyses for the Avenzoariinae and a phylogeny of their charadriiform hosts (compiled from various bird phylogeny hypotheses). The trees could be reconciled by the invoking of 12-13 cospeciation events, 6-7 duplications, 2 host shifts, and 26-29 sorting events. This suggests a high degree of cospeciation.

Animals↗

Enhanced allergic responses to house dust mite by oral exposure to carbaryl in rats.

Epidemiological studies have demonstrated an association between use of carbamate insecticides, including carbaryl, and increased incidence of allergic asthma in farmers. In this study the effect of oral carbaryl exposure on the development of allergic responses to house dust mite (HDM) was examined in female Brown Norway rats. Rats were gavaged for 2 weeks with 0, 2, 10, or 50 mg/kg/day of carbaryl. They were sensitized with a subcutaneous injection of HDM in aluminum hydroxide adjuvant 3 days after the beginning of carbaryl exposure and challenged with antigen via the trachea 1 day after the final carbaryl ingestion. Two days after challenge, antigen-specific cell proliferation in pulmonary lymph nodes was significantly higher in the 50 mg/kg group than in controls, while antigen-specific splenocyte proliferation was decreased in groups dosed with 2, 10, and 50 mg/kg carbaryl. Total protein and lymphocyte number in bronchoalveolar lavage (BAL) fluid were also increased in the 50 mg/kg group. By 7 days after challenge, immune-mediated pulmonary inflammation (eosinophils), antigen-specific immunoglobulin (Ig) E level in serum, and antigen-specific IgE and IgA levels in BAL fluid were significantly elevated in the 50 mg/kg group. No apparent change was observed for lactate dehydrogenase and eosinophil peroxidase in BAL fluid, while the number of BAL macrophages were decreased in groups dosed with 10 and 50 mg/kg carbaryl. The results suggest that carbaryl may cause systemic immune suppression, while enhancing pulmonary allergic responses to house dust mite antigen.

Animals↗

Intraspecific diversity of the Cassava green mite Mononychellus progresivus (Acari: Tetranychidae) using comparisons of mitochondrial and nuclear ribosomal DNA sequences and cross-breeding.

Intraspecific diversity in Cassava Green Mite Mononychellus progresivus Doreste was examined using individuals collected in Benin and the Congo and in Columbia and Brazil. Comparisons were based on mitochondrial and ribosomal DNA sequences and the results of several cross-breeding experiments. Sequence variation was examined in a total of 1139 base pairs (bp) constituting the ITS2 ribosomal DNA (805 bp) and a fragment of the Cytochrome Oxidase I (COI) gene (334 bp). Sequence divergence is low, ranging from 0% to 2.1% for COI and from 0% to 0.4% for ITS2. Inter-strain comparisons have shown that the two African populations appear to be identical. They were similar to the Colombian population while the Brazilian population was clearly different. The data support the hypothesis of a single introduction of the species in the two African populations. Crossing experiments have shown partial hybrid sterility, suggesting a genetic incompatibility consistent with differences detected by sequence data. The results show the usefulness of molecular markers as a tool for determining taxonomic status and dispersion paths in spider mites.

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Transient expression of a Drosophila melanogaster hsp70 promoter/lacZ construct injected into larvae of two species of predatory mites (Acari: Phytoseiidae).

The Drosophila melanogaster heat shock 70 promoter (hsp70) was used to regulate expression of the Escherichia coli beta-galactosidase gene (lacZ) in transiently-transformed predatory mite larvae. A construct containing the hsp70 promoter upstream of the D. melanogaster alcohol dehydrogenase (adh) translational start site and Escherichia coli lacZ gene fusion (adh/lacZ) was injected into larvae of Metaseiulus occidentalis and Amblyseius finlandicus. LacZ expression was compared to expression of a similar construct lacking any upstream regulatory sequence. Expression from the hsp70 promoter was strong and heat shock-dependent in both species. The Drosophila hsp70 promoter therefore appears useful for regulating expression of exogenous DNA in both phytoseiid species and may be broadly applicable in the Phytoseiidae. Furthermore, the lacZ gene is a useful gene for analysis of expression in both species. Larval microinjection provides a method of assessing transient expression and of examining native regulatory sequences in these two phytoseiids and will likely be useful in other phytoseiid mites with only minor modifications.

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Penetration and fate of methomyl and its oxime metabolite in insects and twospotted spider mites.

The penetration and fate of methomyl and its oxime metabolite, methomyl oxime or S-methyl N-hydroxythioacetimidate, were examined in house flies (Musca domestica Linnaeus), face flies (Musca autumnalis De Geer), black cutworm larvae (Agrotis ipsilon (Hufnagel), and twospotted spider mites (Tetranychus urticae Koch). Generally, penetration and metabolism of methomyl-1-14C were rapid in insects and spider mites although differences in rates among the different test organisms were observed. Metabolites from methomyl included CO2, methomyl oxime, and several unknowns. Penetration and metabolism of methomyl oxime-1-14C also were rapid in insects. Metabolites from methomyl oxime included a small amount of CO2 and several unidentified compounds. Organosoluble metabolites from methomyl oxime generally displayed similar chromatographic behavior as those from methomyl. Methomyl was hydrolyzed to its oxime metabolite, which, apparently, was further metabolized to 14CO2 presumably via a Beckmann rearrangement reaction.

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High affinity IgE receptor (Fc epsilon RI) expression on eosinophils infiltrating the lesions and mite patch tested sites in atopic dermatitis.

Expression of the high affinity IgE receptor (Fc epsilon RI) on eosinophils has recently been reported. This led us to evaluate Fc epsilon RI expression on eosinophils in atopic dermatitis (AD). Double immunofluorescence stainings with an anti-Fc epsilon RI monoclonal antibody (mAb) and a polyclonal antieosinophil cationic protein (ECP) antibody were performed on lesional biopsy specimens from patients with AD and from patients with bullous pemphigoid (BP) as controls. In AD and BP lesions, 77% and 70% of eosinophils expressed Fc epsilon RI, respectively. However, the intensity of Fc epsilon RI staining in AD was much stronger than in BP, suggesting upregulation of Fc epsilon RI expression on eosinophils in AD. In addition, the eosinophils infiltrating AD lesions were stained strongly with anti-CD23 mAb and anti-IgE antibody. At the sites of mite patch testing in AD, Fc epsilon RI-, CD23- and IgE-positive eosinophils were observed to the same degree as in the lesions, and a considerable number of mite antigen-bearing eosinophils were detected. Fc epsilon RI and CD23 were both upregulated on the skin-infiltrating eosinophils in AD and bound IgE molecules.

Adult↗

Differentiation of mite species by using cellulose acetate and equilibrium polyacrylamide gel electrophoresis of isoenzymes.

The isoenzymes of various species of medically and economically important mites were studied using cellulose acetate and equilibrium polyacrylamide gel electrophoresis. Interspecific differences in isoenzymes were found, which were species-specific. Intraspecific differences in isoenzymes were also found when individual mites were examined. The efficiency of each technique, and their use in various fields of acarology, are discussed. In addition, possible phylogenetic relationships as revealed by these techniques are suggested.

Animals↗

Ultrastructure of the reproductive system of the house dust mites Dermatophagoides farinae and D. pteronyssinus (Acari, Pyroglyphidae) with remarks on spermatogenesis and oogenesis.

Several parts of the reproductive system of both sexes of Dermatophagoides farinae and D. pteronyssinus are investigated and compared by light-, scanning electron-, and transmission electron microscopy. New techniques have been employed for scanning of the internal structures of these mites. The male reproductive system consists of unpaired testis, paired vasa deferentia, an accessory gland, ejaculatory duct, and copulatory organ. The female reproductive system consists of bursa copulatrix, ductus bursae, receptaculum seminis, paired ducti receptaculi, ovaries, oviducts, one chorion gland, ovipositor, and oviporus. Testis as well as ovaries are characterized by syncytia of nutritional function. The specifics of spermatogenesis are discussed in connection with sperm transfer. Similarities between the construction of the ovaries and oogenesis in astigmatic mites and telotrophic meroistic ovaries in insects are discussed.

Animals↗

Molecular polymorphisms of house dust mite allergens.

Previous studies from this laboratory have described the primary amino acid sequences of the group I and group II allergens from Dermatophagoides pteronyssinus and D. farinae. This report concentrates on polymorphisms of allergens within a species. Firstly, four cDNA clones of Der fII produced by polymerase chain reaction have been sequenced and are compared to the sequences published previously by ourselves and others. Although the sequences come from different sources, Australia and Japan, the overriding conclusion is one of similarity, with only two possible non-conservative changes in the six sequences. The nucleotides were also very conserved including the 3' untranslated regions, although some non-coding differences could be found which may provide a genetic marker. Experiments are reported to help define the group III D. pteronyssinus allergens. Previous studies have characterised the group III of D. farinae as a Mr 29-kDa molecule which can be defined by monoclonal antibodies. A Mr 17-kDa molecule of D. pteronyssinus has been reported with an almost identical N-terminal sequence. Here it is described that Der fIII isolated from different preparations of spent mite media by affinity chromatography have predominantly Mr 32-, 28- and 21-kDa forms which vary in degree from batch to batch. 83% of adults and 38% of children react with the preparation by radioimmune dot-blot. The difference between the children and adults is statistically significant and reactivity can be to at least the 32- and 28-kDa form. Antisera produced in mice against the Der fIII react to D. pteronyssinus mite extract by Western blotting primarily to a 32-kDa moiety, but also 28- and 21-kDa forms in some extracts.

Allergens↗

Control of house-dust mites (Pyroglyphidae) with home disinfectants.

Chemical disinfectants and biocidal preparations used in households were tested in the laboratory for their ability to kill the house-dust mite Dermatophagoides farinae. Batches of ten specimens were soaked in aqueous solutions or suspensions containing 0.0, 0.1, 0.3, 1.0, 3.0 and 10.0% (by volume) of the test preparations. Direct effect was tested without food. Population effect was tested with food added. The results showed a high mortality with all preparations except for a regular carpet cleaner (containing detergents) and natamycin (a fungicide). Nevertheless, not all tested preparations are practical in the home environment. Best results in homes were obtained with a carpet cleaning solution which incorporates an acaricide (benzylbenzoate). This particular preparation has an outstanding acaricidal efficacy and can easily and routinely be used by the householder. The degree of cleanliness in the household is a measure of the number of house-dust mites and their allergens.

Allergens↗

Possible members of the potyvirus group transmitted by mites or whiteflies share epitopes with aphid-transmitted definitive members of the group.

There are at least ten viruses identified in the literature that resemble definitive potyviruses in having flexuous filamentous particles and inducing the formation of "pinwheel" cytoplasmic inclusions in infected cells but that are transmitted by eriophyid mites, whiteflies or soil fungi and not by aphids, the vectors of the definitive potyviruses. The taxonomic status of these viruses is uncertain at present. Using a broadly cross-reactive antiserum raised against the dissociated coat protein core (residues 68-285) of a definitive potyvirus (Johnsongrass mosaic virus), we have shown that wheat streak mosaic virus which is transmitted by mite and sweet potato mild mottle virus which is transmitted by whitefly have coat proteins that share epitopes with definitive potyviruses. This finding further supports their classification as definitive members of the potyvirus group. The cross-reactive antiserum used here had been shown previously to react with coat proteins of fifteen different definitive potyviruses. The antiserum did not react with coat proteins of potexviruses and tobamoviruses.

Animals↗

Mitochondrial DNA from a spider mite: isolation, restriction map and partial sequence of the cytochrome oxidase subunit I gene.

Mitochondrial (mt) DNA of the phytophagous mite Tetranychus urticae was purified and a restriction map was constructed. The 12.5 kb long genome is the shortest animal mtDNA known. A 564 bp clone comprising part of the gene for cytochrome oxidase subunit I was sequenced. As has been found in insects, the mitochondrial sequences of mites are extremely A+T rich (75% on average, 96.5% at the third codon position).

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Dopamine and octopamine in whole body extracts of the bulb mite.

Using high performance liquid chromatography with electrochemical detection, whole body extracts of the bulb mite, Rhizoglyphus echinopus (Fumouze and Robin), were found to contain the biogenic amines dopamine and octopamine at concentrations of 4.3 +/- 0.6 and 2.3 +/- 1.4 ng g-1 wet weight, respectively. Adrenaline, noradrenaline, tyramine, N-methyldopamine, N-acetyldopamine, and 5-hydroxytryptamine, if present, were below the limits of detectability. This is the initial demonstration of the presence of octopamine in a mite species.

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Surface skin lipids of birds--a proper host kairomone and feeding inducer in the poultry red mite, Dermanyssus gallinae.

A factor in bird skin responsible for D. gallinae feeding on its natural hosts was investigated. Both skin and plumage of chickens contained substance(s) attractive to mites which was easily dissolved in benzene derivates, amyl acetate or ethyl acetate, thermostabile up to 100 degrees C, nonvolatile, alkaline hydrolysable, and susceptible to oxidation. Components of surface skin lipids were ascertained to be the host-markers when using chromatographic separation and consecutive in vitro feeding technique for testing the isolates. Purified specific fowl diol esters of fatty acids prepared from the secretion of uropygial (preen) glands of hens were at least as effective a feeding stimulant for the mites as the natural extract of surface lipids of birds.

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Bifurcation and stability analysis of a temperature-dependent mite predator-prey interaction model incorporating a prey refuge.

The non-linear behavior of a differential equations-based predator-prey model, incorporating a spatial refuge protecting a constant proportion of prey and wit temperature-dependent parameters chosen appropriately for a mite interaction on fruit trees, is examined using the numerical bifurcation code AUTO 86. The most significant result of this analysis is the existence of a temperature interval in which increasing the amount of refuge dynamically destabilizes the system; and on part of this interval the interaction is less likely to persist in that predator and prey minimum population densities are lower than when no refuge is available. It is also shown that increasing the amount of refuge can lead to population outbreaks due to the presence of multiple stable states. The ecological implications of a refuge are discussed with respect to the biological control of mite pests.

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