Modelling and Bioinformatics Analysis of the Dimeric Structure of House Dust Mite Allergens from Families 5 and 21: Der f 5 Could Dimerize as Der p 5

Abstract

Allergy represents an increasing thread to public health in both developed and emerging countries and the dust mites Dermatophagoides pteronyssinus (Der p), Blomia tropicalis (Blo t), Dermatophagoides farinae (Der f), Lepidoglyphus destructor (Lep d) and Suidasia medanensis (Sui m) strongly contribute to this problem. Their allergens are classified in several families among which families 5 and 21 which are the subject of this work. Indeed, their biological function as well as the mechanism or epitopes by which they are contributing to the allergic response remain unknown and their tridimensional structures have not been resolved experimentally except for Blo t 5 and Der p 5. Blo t 5 is a monomeric three helical bundle, whereas Der p 5 shows a three helical bundle with a kinked N-terminal helix that assembles in an entangled dimeric structure with a large hydrophobic cavity. This cavity could be involved in the binding of hydrophobic ligands, which in turn could be responsible for the shift of the immune response from tolerance to allergic inflammation. We used molecular modelling approaches to bring out if other house dust mite allergens of families 5 and 21 (Der f 5, Sui m 5, Lep d 5, Der p 21 and Der f 21) could dimerize and form a large cavity in the same way as Der p 5. Monomeric models were first performed with MODELLER using the experimental structures of Der p 5 and Blo t 5 as templates. The ClusPro server processed the selected monomers in order to assess their capacity to form dimeric structures with a positive result for Der p 5 and Der f 5 only. The other allergens (Blo t 5, Sui m 5, Lep d 5, Der p 21 and Der f 21) did not present such a propensity. Moreover, we identified mutations that should destabilize and/or prevent the formation of the Der p 5 dimeric structure. The production of these mutated proteins could help us to understand the role of the dimerization process in the allergic response induced by Der p 5, and if Der p 5 and Der f 5 behave similarly.     

贵州梵净山自然保护区土壤甲螨群落的季节动态

2001年7月至2002年4月在不同的季节,对梵净山自然保护区属级水平上土壤甲螨结构的季节变化进行了调查.结果显示,土壤甲螨多度的四季变化为春季＞冬季＞秋季＞夏季.土壤甲螨多样性的四季变化为秋季＞夏季＞冬季＞春季.不同的季节土壤甲螨群落组成不同,但小奥甲螨属、小盾珠甲螨属、木单翼甲螨属3个属在每个季节均为绝对优势属.土壤甲螨主要分布在表层,但从夏季到冬季,土壤甲螨从表层向深层迁徙.     

Tunicamycin Inhibits Biosynthesis of Acid Mucopolysaccharides in Cultures of Chick Embryo Fibroblasts

Enterobacter cloacae KY 3074 grown in a medium containing xanthine, hypoxanthine, guanine, or their nucleosides and nucleotides produced xanthine oxidase. The purified enzyme preparation showed a major protein band and a few minor bands in acrylamide gel electrophoresis. Molecular oxygen was the most effective electron acceptor. Ferricyanide and 2,6-dichlorophenolindophenol also served as electron acceptors, but NAD and NADP did not. Xanthine and hypoxanthine were good substrates, and guanine was also an effective substrate. The activity was inhibited by Ag²⁺, Cu²⁺, PCMB, and ascorbate. The spectrum of the Enterobacter enzyme resembled that of some known xanthine oxidizing enzymes, and this suggests a similarity in the prosthetic groups of these enzymes. The molecular weight of the native enzyme and subunit was 128,000 and 69,000, respectively.     

Genes important for survival or reproduction in Varroa destructor identified by RNAi

The Varroa mite,(Varroa destructor),is the worst threat to honey bee health worldwide.To explore the possibility of using RNA interference to control this pest, we determined the effects of knocking down various genes on Varroa mite survival and reproduction.Double-stranded RNA (dsRNA)of six candidate genes (Da,Pros26S,RpL8, RpL11,RpPO and RpS13)were synthesized and each injected into Varroa mites,then mite survival and reproduction were assessed.Injection of dsRNA for Da (Daughterless)and Pros26S (Gene for proteasome 26S subunit adenosine triphosphatase)caused a significant reduction in mite survival,with 3.57%±1.94% and 30.03%±11.43% mites surviving at 72 h post-inj ection (hpi),respectively.Control mites injected with green fluorescent protein (GFP)-dsRNA showed survival rates of 81.95%±5.03% and 82.36 ±2.81%,respectively. Injections of dsRNA for four other genes (RpL8,RpL11,RpPO and RpS13)did not affect survival significantly,enabling us to assess their effect on Varroa mite reproduction.The number of female offspring per mite was significantly reduced for mites injected with dsRNA of each of these four genes compared to their GFP-dsRNA controls.Knockdown of the target genes was verified by real-time polymerase chain reaction for two genes important for reproduction (RpL8,RpL11)and one gene important for survival (Pros26S). In conclusion,through RNA interference,we have discovered two genes important for mite survival and four genes important for mite reproduction.These genes could be explored as possible targets for the control of Varroa destructor in the future.     

A new microinvertebrate with features of mites and tardigrades in Dominican amber

From time to time, small, fragile, previously unknown fossil invertebrates are found in specialized habitats. Occasionally, as in the present case, a fragment of the original habitat that existed millions of years ago is also preserved. The present article describes a previously unknown microinvertebrate in Dominican amber that cannot be placed in any group of extant invertebrates. Placed in a new family, genus, and species, the fossil shares characters with both tardigrades and mites, but clearly belongs to neither group. The several hundred fossil individuals preserved in the amber shared their moist, warm habitat with pseudoscorpions, nematodes, fungi, and protozoa. The large number of fossils provided additional evidence of their biology, including their reproductive behavior, developmental stages, and food. While there is no extant group that can accommodate these fossils, and we have no knowledge of any extant descendants, this discovery shows that unique lineages of minute invertebrates were surviving in the mid‐Tertiary.     

Phylogeography of lethal male fighting in a social spider mite

When males fight for access to females, such conflict rarely escalates into lethal fight because the risks and costs involved, that is, severe injury or death, are too high. The social spider mite, Stigmaeopsis miscanthi, does exhibit lethal male fights, and this male–male aggressiveness varies among populations. To understand the evolution of lethal fighting, we investigated aggressiveness in 42 populations and phylogenetic relationships in 47 populations along the Japanese archipelago. By analysis of the male weapon morph, a proxy for aggressiveness, we confirmed the existence of a mildly aggressive (ML) form, besides the low aggression (LW) and high aggression (HG) forms reported earlier. To evaluate demographic history of these three forms, we employed the approximate Bayesian computation approach using mtCOI sequences and taking into consideration the postlast glacial expansion history of the host plant, Miscanthus sinensis. As results, hierarchical split models are more likely to explain the observed genetic pattern than admixture models, and the ML form in the subtropical region was considered the ancestral group. The inferred demographic history was consistent with the one reconstructed for the host plant in a previous study. The LW form was split from the ML form during the last glacial period (20,000–40,000 years BP), and subsequently, the HG form was split from the ML form at the end of or after the last glacial period (5,494–10,988 years BP). The results also suggest that the mite invaded Japan more than once, resulting in the present parapatric distribution of LW and HG forms in eastern Japan.     

Effect of coupled reduced irrigation and nitrogen fertilizer on soil mite community composition in a wheat field

Groundwater and nitrogen fertilizer overuse severely threatens crop productions; thus, current ecological agriculture requires low irrigation and nitrogen fertilizer inputs. The effects of combined reduced irrigation and nitrogen fertilizer addition on soil organism (e.g., mite) community and biodiversity remain poorly understood. We analyzed soil mite community composition, wheat grain yield, and soil characteristics in a 10‐year manipulation experiment with two levels of irrigation (reduced and conventional irrigation) and five nitrogen fertilizer levels (0, 70, 140, 210, and 280 kg N/ha). Reduced irrigation (20% reduction, from 280 to 220 mm) and nitrogen fertilizer (25% reduction, from 280 to 210 kg N/ha) addition did not significantly influence soil mite community and wheat yield. The relative abundances of fungivores and predators showed negative quadratic relationships with wheat yield, while that of plant parasites showed a positive relationship. The relationships between soil mite trophic groups and wheat yield revealed that we can evaluate the impacts of reduced irrigation and nitrogen fertilizer addition from the perspective of soil fauna. Soil mite community composition was altered by soil abiotic factors prior to reduced irrigation and nitrogen fertilizer addition. Overall, moderate reductions of irrigation and nitrogen fertilizer may not threaten to soil mite community and diversity or decrease crop production; in contrast, such reductions will benefit mite community development and the sustainable agriculture.     

Life table characteristics of Macrolophus caliginosus preying upon Tetranychus urticae

The life table characteristics of the polyphagous mirid Macrolophus caliginosus Wagner (Heteroptera: Miridae) preying on various stages of Tetranychus urticae Koch (Acari: Tetranychidae) with tomato as host plant were described at 22 °C. The following average parameters were obtained: Female longevity: 28.7 days; fecundity: 0.7 eggs/female/day; egg mortality: 2.6%; pre-oviposition period: 5.5 days; oviposition period: 18.1 days; post-oviposition period: 3.2 days; juvenile development time: 26.8 days; juvenile mortality: 34.9%; and sex ratio (/(+): 0.46. Life table parameters were estimated as net reproduction rate (R ₀): 6.15; intrinsic rate of increase (r _m): 0.031 day^–1; finite rate of increase (): 1.032; mean generation time (T _c): 58.17 days; and doubling time (T ₂) 22.2 days. The parameters obtained were in accordance with those reported for M. caliginosus fed on another mite species (T. turkestani Ugarov & Nikolski (Acari: Tetranychidae)). However, compared to the performance of M. caliginosus fed on common glasshouse insect pests, a diet consisting of only mites appeared to be inferior. However, being a voracious predator, M. caliginosus may be a valuable addition to existing methods of mite control.     

Proteolytic activities in body and faecal extracts of the storage mite, Acarus farris

Trypsin, chymotrypsin, cathepsins B and D, aminopeptidase and carboxypeptidases A and B were detected in body extracts of the storage mite Acarus farris (Oudemans) (Astigmata: Acaridae). Faeces-enriched medium exhibited higher (10-50-fold) specific protease activity rates than those measured with mite body extracts for trypsin, chymotrypsin and carboxypeptidases A and B, suggesting that they are involved in mite digestion. However, the activity of cathepsin B was only three-fold higher in faecal than in body extracts, indicating that its presence in the lumen of the digestive tract is low compared to that of serine proteases. The activity of aminopeptidases was higher in mite bodies, indicating that they might be membrane bound. Cathepsin D activity was only detected in body extracts, indicating that this enzyme is not a digestive protease in this species. Zymograms resolved three major bands of gelatinolytic activity, but at least one protease form was only present in body extracts. Protease inhibitors of different specificity were tested in vivo to establish their potential as control agents. The development of A. farris was significantly retarded when the immature stages were fed on artificial diet containing inhibitors of serine and cysteine proteases and aminopeptidases, whereas no such effect was found with inhibitors of aspartyl proteases and carboxypeptidases. Interestingly, the most significant effects on A. farris occurred when a combination of inhibitors targeting different enzyme classes was supplied mixed in the diet, suggesting a synergistic toxicity. Several plant lectins were also tested, but only wheat germ agglutinin and concanavalin-A affected development.