Parasitism and venom of ectoparasitoid Scleroderma guani impairs host cellular immunity.

Venom is a prominently maternal virulent factor utilized by parasitoids to overcome hosts immune defense. With respect to roles of this toxic mixture involved in manipulating hosts immunity, great interest has been mostly restricted to Ichneumonoidea parasitoids associated with polydnavirus (PDV), of which venom is usually considered as a helper component to enhance the role of PDV, and limited Chalcidoidea species. In contrast, little information is available in other parasitoids, especially ectoparasitic species not carrying PDV. The ectoparasitoid Scleroderma guani injects venom into its host, Tenebrio molitor, implying its venom was involved in suppression of hosts immune response for successful parasitism. Thus, we investigated the effects of parasitism and venom of this parasitoid on counteracting the cellular immunity of its host by examining changes of hemocyte counts, and hemocyte spreading and encapsulation ability. Total hemocyte counts were elevated in parasitized and venom-injected pupae. The spreading behavior of both granulocytes and plasmatocytes was impaired by parasitization and venom. High concentration of venom led to more severely increased hemocyte counts and suppression of hemocyte spreading. The ability of hemocyte encapsulation was inhibited by venom in vitro. In addition to immediate effects observed, venom showed persistent interference in hosts cellular immunity. These results indicate that venom alone from S. guani plays a pivotal role in blocking hosts cellular immune response, serving as a regulator that guarantees the successful development of its progenies. The findings provide a foundation for further investigation of the underlying mechanisms in immune inhibitory action of S. guani venom.

Phytotoxic and Antimicrobial Terrein Derivatives and Butenolides Isolated from the Endophytic Fungus Aspergillus terreus HT5.

Two new terrein derivatives, aspergilethers A and B (1 and 2), two known analogues (3 and 4), and three known butenolides (5-7) were isolated from the endophyte Aspergillus terreus HT5. Their structures were determined by spectroscopic analysis and ECD and NMR calculations. Interestingly, 1 and 2 had unpresented medium aliphatic side chains in terrein derivatives, with different absolute configurations at C-7, which was very scarce. (+)-Terrein (3) exhibited potent postemergence phytotoxicity toward Amaranthaceae, Portulacaceae, and Fabaceae, with MIC values of 250-1000 µg/mL. Transcriptome analysis and qRT-PCR suggested that (+)-terrein induced the transcriptional expression of aging-related genes to accelerate organ senescence and stimulated plant detoxification response. The conjugated system between keto carbonyl and double bonds in the cyclopentenone ring and side chain, and the configurations of C-2 and C-3, played critical roles in the phytotoxicity of terrein derivatives. Meanwhile, 3 was first reported to display moderate antioomycetes activity toward Phytophthora nicotiana.

Phytotoxic Azaphilones From the Mangrove-Derived Fungus Penicillium sclerotiorum HY5.

Mangrove is a unique marine ecosystem growing in the intertidal zone of tropical and subtropical coast, with the characteristics of hypoxia tolerance, high salinity, and high humidity. In order to discover novel leading compounds with potent phytotoxicity, seven pairs of azaphilones E/Z isomers, isochromophilone H (1a/1b), sclerotiorins A and B (2a/2b and 3a/3b), ochlephilone (4a/4b), isochromophilone IV (5a/5b), isochromophilone J (6a/6b), and isochromophilone I (7a/7b), were isolated from the culture broth of the mangrove-derived fungus, the Penicillium sclerotiorum HY5, by various chromatographic methods. Among them, 1a, 1b, 2a, 3a, 4a, 5a, 6a, and 6b were new compounds. Their chemical structures and absolute configurations were elucidated based on high resolution electrospray ionization mass spectroscopy (HRESIMS), 1D/2D nuclear magnetic resonance (NMR) spectroscopic analysis, and comparisons of electronic circular dichroism (ECD) data. Compounds 3, 4, and 7 exhibited potent phytotoxicity against the growth of radicle and plumule on Amaranthus retroflexus L., with EC50 values ranging from 234.87 to 320.84 µM, compared to the positive control glufosinate-ammonium, with EC50 values of 555.11 µM for radicle, and 656.04 µM for plumule. Compounds 4 and 7 also showed inhibitory effects on the growth of velvetleaf (Abutilon theophrasti Medikus), with EC50 values ranging from 768.97 to 1,201.52 µM. This study provides new leading compounds for the research and development of marine-derived bioherbicides.

Observation of the fine structure of antennal sensilla of the stink bug, Eocanthecona furcellata (Hemiptera: Pentatomidae).

In order to explore the mechanism underlying chemosensation in Eocanthecona furcellata, the external morphology of its antennae and the type, quantity, distribution and ultrastructure of the sensilla were observed on both sexes of adults and 5th-instar nymphs using scanning electron microscopy. The results showed that the antennae of E. furcellata consisted of three parts: scape, pedicel and flagellum. There were five types of sensilla on the antennae, which included sensilla trichoidea (ST), sensilla chaetica (SCh), sensilla coeloconica (SCo), sensilla basiconica (SB) and squamifornia denticles (SD). Further, there were 4 subtypes of ST and SB and 2 subtypes of SCo and SCh. The number of sensilla on nymphs was significantly lower than that on adults. The antennae of adults showed sexual dimorphism, as the number of sensilla on female adults was higher than that on male adults. SB4 was found only on females and SCo2 was found only on males. These inter-sexual differences may be related to chemoreception of sex pheromone and chemical predation location. The morphology and putative functions of each sensilla were compared and discussed. These results provide a reference for further study of the behavioral biology, chemical ecology and electrophysiology of insects, and also provides a scientific basis for new ways of biological control.

Ultrastructural alterations in midgut and Malpighian tubules of Boettcherisca peregrina exposure to cadmium and copper.

The effects of Cu and Cd at their at their low concentrations (80microg/g diet) on the morphology and ultrastructure of the midgut and Malpighian tubules of Boettcherisca peregrina larvae were observed by light and transmission electron microscopy. After exposure to both metals, the midgut got darker, shorter, and thicker than in control, and many strumae occurred on the surface of the midgut. Similarly, Malpighian tubules got shorter and thinner. Ultrastructural alterations in the midgut included mitochondrial condensation, swelling, and lysis. The rough endoplasmic reticulum (rER) showed dilation and vesiculation. The microvilli were shortened and disorganized. The stored glycogens increased and many mineral spherites appeared along with lipid droplets decreased. Ultrastructural alterations observed in the Malpighian tubules included rER vesiculation and mitochondria swelling with loss of cristae. Shortened and disordered microvilli, increased numbers of large hydropic vacuoles, and mineral spherites were also observed.

Five monocyclic pyridinium derivative based halo-argentate/cuprate hybrids or iodide salts: influence of composition on photochromic behaviors.

The rational choice of an electron acceptor was proved to be an effective strategy for the development of novel electron transfer (ET) photochromic iodides, but the types and amounts of reported electron acceptors are relatively limited so far, especially for monocyclic aromatic molecules. Herein, using monocyclic pyridinium derivatives (N-protonation-4-carboxypyridinium/N-protonation-4-carbamoylpyridinium/1-methyl-4-(carbomethoxy)pyridinium) as structural directing agents and electron acceptors, five new electron donor-acceptor-based halo-argentate/cuprate hybrids or iodide salts have been synthesized, including [HINA][Ag4I5] (1), [HINAM]I (2), [HINAM]I·0.5(I2) (3), [MCMP][Ag2Br3] (4) and [MCMP][Cu2I3] (5). Noteworthily, compounds 1-3 exhibit interesting photochromic behaviours, while compounds 4 and 5 are non-photochromic. Finally, the possible chromic mechanisms and influencing factors for the title compounds were also discussed.

Astronaut-centered philosophy for designing manned space system.

Astronaut-centered design philosophy is a new concept suggested by the authors for manned space system design. It stems from human-centered design philosophy. Human-centered design means that human role is regarded as important basis and foundation for system design. At the beginning, the engineers used to adopt technology-centered philosophy for designing complex system, but much practice proved that the technology-centered design philosophy won't work, resulting in lower system safety and performance. So it has been currently replaced by human-centered philosophy. As examples, the principles of human-centered automation of the International Civil Aviation Organization and NASA JSC's Human-rating Requirements were introduced. At last, the astronaut-centered design philosophy and its requirements were put forward by the authors. These requirements consist of: general requirements, man-machine interaction requirements, man-environment interaction requirements and interpersonal relationship requirements.

Parasitization by Scleroderma guani influences protein expression in Tenebrio molitor pupae.

Ectoparasitoid wasps deposit their eggs onto the surface and inject venom into their hosts. Venoms are chemically complex and they exert substantial impact on hosts, including permanent or temporary paralysis and developmental arrest. These visible venom effects are due to changes in expression of genes encoding physiologically relevant proteins. While the influence of parasitization on gene expression in several lepidopterans has been reported, the molecular details of parasitoid/beetle relationships remain mostly unknown. This shortcoming led us to pose the hypothesis that envenomation by the ectoparasitic ant-like bethylid wasp Scleroderma guani leads to changes in protein expression in the yellow mealworm beetle Tenebrio molitor. We tested our hypothesis by comparing the proteomes of non-parasitized and parasitized host pupae using iTRAQ-based proteomics. We identified 41 proteins that were differentially expressed (32↑- and 9↓-regulated) in parasitized pupae. We assigned these proteins to functional categories, including immunity, stress and detoxification, energy metabolism, development, cytoskeleton, signaling and others. We recorded parallel changes in mRNA levels and protein abundance in 14 selected proteins following parasitization. Our findings support our hypothesis by documenting changes in protein expression in parasitized hosts.

Heat shock protein genes (hsp20, hsp75 and hsp90) from Pieris rapae: molecular cloning and transcription in response to parasitization by Pteromalus puparum.

Most molecular work on the roles of heat shock proteins (hsps) in host-parasite interaction has focused on vertebrates, rather than invertebrates. Here the full length complementary DNA (cDNA) sequences of three hsp genes (hsp20, hsp75 and hsp90) were amplified from Pieris rapae, and their transcriptional responsiveness to parasitization by the endoparasitic wasp Pteromalus puparum were investigated. The cDNA sequence analysis of hsp20, hsp75 and hsp90 revealed open reading frames of 531, 2 328 and 2 157 bp in length, which encode proteins with calculated molecular weights of 19.5, 75.48 and 82.7 kDa, respectively. The comparison of amino acid sequences showed that P. rapae hsp20 shared highly divergent homology to that of other insects, while hsp75 and hsp90 showed high homology to their counterparts of other species. The expression analysis indicated that these three genes were influenced in response to parasitization by P. puparum. The hsp20 transcripts in parasitized pupae were higher compared to non-parasitized pupae. The expression of hsp75 and hsp90 were down-regulated following parasitization. The results indicate that hsps are involved in host-parasitoid interactions.

Transcriptomic immune response of Tenebrio molitor pupae to parasitization by Scleroderma guani.

BACKGROUND: Host and parasitoid interaction is one of the most fascinating relationships of insects, which is currently receiving an increasing interest. Understanding the mechanisms evolved by the parasitoids to evade or suppress the host immune system is important for dissecting this interaction, while it was still poorly known. In order to gain insight into the immune response of Tenebrio molitor to parasitization by Scleroderma guani, the transcriptome of T. molitor pupae was sequenced with focus on immune-related gene, and the non-parasitized and parasitized T. molitor pupae were analyzed by digital gene expression (DGE) analysis with special emphasis on parasitoid-induced immune-related genes using Illumina sequencing. METHODOLOGY/PRINCIPAL FINDINGS: In a single run, 264,698 raw reads were obtained. De novo assembly generated 71,514 unigenes with mean length of 424 bp. Of those unigenes, 37,373 (52.26%) showed similarity to the known proteins in the NCBI nr database. Via analysis of the transcriptome data in depth, 430 unigenes related to immunity were identified. DGE analysis revealed that parasitization by S. guani had considerable impacts on the transcriptome profile of T. molitor pupae, as indicated by the significant up- or down-regulation of 3,431 parasitism-responsive transcripts. The expression of a total of 74 unigenes involved in immune response of T. molitor was significantly altered after parasitization. CONCLUSIONS/SIGNIFICANCE: obtained T. molitor transcriptome, in addition to establishing a fundamental resource for further research on functional genomics, has allowed the discovery of a large group of immune genes that might provide a meaningful framework to better understand the immune response in this species and other beetles. The DGE profiling data provides comprehensive T. molitor immune gene expression information at the transcriptional level following parasitization, and sheds valuable light on the molecular understanding of the host-parasitoid interaction.

Heterometallic mixed-valence copper(I,II) cyanides that were tuned by using the chelate effect: discovery of famous Cairo pentagonal tiling and unprecedented (3,4)-connected {8(3)}2{8(6)} topological 3D net.

By using environmentally friendly [Ni(CN)4](2-) as a cyanide source, three new heterometallic cyano-bridged mixed-valence Cu(I)/Cu(II) coordination polymers with three different electronic configurations (d(8)-d(10)), that is, [Cu2Ni(CN)5(H2O)3] (1), [Cu2Ni(CN)5(pn)H2O] (2), and [Cu3Ni(CN)6(pn)2] (3, pn = 1,2-propane diamine) have been synthesized by gradually increasing the amount of pn. Compound 1, which was hydrothermally synthesized in the absence of pn ligand, exhibits the famous 2D Cairo pentagonal tiling, in which the Cu(I), Cu(II), and Ni(II) atoms act as trigonal, T-shaped, and square-planar nodes, respectively. Notably, there are three water molecules located at the meridianal positions of the octahedrally coordinated Cu(II) atom in compound 1. A similar reaction, except for the addition of a small amount of pn, generated a similar Cairo pentagonal tiling layer in which two of the water molecules that were located at the meridianal positions of the octahedrally coordinated Cu(II) atom were replaced by a chelating pn group. Another similar hydrothermal reaction, with the addition of a larger amount of pn, yielded compound 3, which showed a related two-fold-interpenetrated (3,4)-connected 3D framework with an unprecedented {8(3)}2{8(6)} topology in which the Cu(II) atom was chelated by two pn groups. These structural changes between compounds 1-3 can be explained by the chelating effect of the pn group. The replacement of two meridianally coordinated water molecules on the octahedral Cu(II) atom in compound 1 by a pn group gives compound 2, which shows similar Cairo tiling, and a further increase in the amount of pn results in the formation of the [Cu(NC)2(pn)2] unit and the two-fold-interpenetrated 3D framework of compound 3. The mixed-valence properties of compounds 1, 2, and 3 were confirmed by variable-temperature magnetic-susceptibility measurements.

High-throughput genotyping of single-nucleotide polymorphisms in ace-1 gene of mosquitoes using MALDI-TOF mass spectrometry.

Acetylcholinesterase (AChE) plays a vital role in the nervous system of insects and other animal species and serves as the target for many chemical agents such as organophosphate and carbamate insecticides. The mosquito, Culex pipiens complex, a vector of human disease, has evolved to be resistant to insecticides by a limited number of amino acid substitutions in AChE1, which is encoded by the ace-1 gene. The aims of this study are to identify single nucleotide polymorphism (SNP) sites in the ace-1 gene of the C. pipiens complex and explore an economical high-throughput method to differentiate the genotypes of these sites in mosquitoes collected in the field. We identified 22 SNP sites in exon regions of the ace-1 gene. Four of them led to non-synonymous mutations, that is, Y163C, G247S, C677S and T682A. We used matrix-assisted laser desorption ionization - time-of-flight mass spectrometry for genotyping at these four sites and another site F416V, which was relevant to insecticide resistance, in 150 mosquitoes collected from 15 field populations. We were able to synchronize analysis of the five SNP sites in each well of a 384-well plate for each individual mosquito, thus decreasing the cost to one-fifth of the routine analysis. Heterozygous genotypes at Y163C and G247S sites were observed in one mosquito. The possible influence of the five SNP sites on the activity or function of the enzyme is discussed based on the predicted tertiary structure of the enzyme.

Neopierisoids A and B, two new chlorinated 3,4-seco-grayanane diterpenoids with antifeedant activity from flowers of Pieris japonica.

Two new chlorinated multiacylated 3,4-seco-grayanane diterpenoids, neopierisoids A and B (1 and 2), were isolated from flowers of the poisonous plant Pieris japonica and were identified from spectroscopic analysis and X-ray diffraction data. Both compounds showed obvious antifeedant activity against Pieris brassicae with an EC50 of 10.07 µg/cm(2) for 1 and 5.33 µg/cm(2) for 2, indications of toxic properties. Chlorinated 3,4-seco-grayanane diterpenoids in P. japonica may play a defensive role against herbivores.

Prophenoloxidase from Pieris rapae: gene cloning, activity, and transcription in response to venom/calyx fluid from the endoparasitoid wasp Cotesia glomerata.

Prophenoloxidase (PPO) plays an important role in melanization, necessary for defense against intruding parasitoids. Parasitoids have evolved to inject maternal virulence factors into the host hemocoel to suppress hemolymph melanization for the successful development of their progeny. In this study, the full-length complementary DNA (cDNA) of a Pieris rapae PPO was cloned. Its cDNA contained a 2 076-base pair (bp) open reading frame (ORF) encoding 691 amino acids (aa). Two putative copper-binding sites, a proteolytic activation site, three conserved hemocyanin domains, and a thiol ester motif were found in the deduced amino acid sequence. According to both multiple alignment and phylogenetic analysis, P. rapae PPO gene cloned here is a member of the lepidopteran PPO-2 family. Injection of Cotesia glomerata venom or calyx fluid resulted in reduction of P. rapae hemolymph phenoloxidase activity, demonstrating the ability to inhibit the host's melanization. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) showed that transcripts of P. rapae PPO-2 in the haemocytes from larvae had not significantly changed following venom injection, suggesting that the regulation of PPO messenger RNA (mRNA) expression by venom was not employed by C. glomerata to cause failure of melanization in parasitized host. While decreased P. rapae PPO-2 gene expression was observed in the haemocytes after calyx fluid injection, no detectable transcriptional change was induced by parasitization, indicating that transcriptional down-regulation of PPO by calyx fluid might play a minor role involved in inhibiting the host's melanization.

A hybrid cobalt hydroxyacetate magnet: ionothermal synthesis, 3-D Co-O-Co connectivity and spin glass behavior.

An ionothermal reaction has generated a cobalt hydroxyacetate magnet that contains a three-dimensional Co-O-Co framework consisting of vertex-sharing octahedral chains bridged by another octahedra via edge-sharing and shows spin glass behavior.

[The preliminary study of three-dimensional simulation of the craniofacial system].

OBJECTIVE: To look for the way of three-dimensional simulation of the craniofacial system. METHODS: A three-dimensional laser scanner was used for gypsum models digitization and computed tomography scans was employed for skull reconstruction, then the data of teeth and temporomandibular joint were picked up and integrated. The ARCUS sigma system was used to record spatial mandibular movements. The data of both digital reconstruction and spatial movements were transferred into one coordinate system. The software for three-dimensional simulation was programmed. RESULTS: The preliminary program could be used to analyze static and dynamic occlusion and gnathic relations, to check the contact points and to show from various visual angles and slices. The occlusal plane, curves, and helical axis were initially defined and displayed. CONCLUSIONS: Using available instruments and methods, we developed the primary edition for three-dimensional simulation of the craniofacial system. However, it is far from a mature system and there is still plenty of work to be done.