Search

The brown planthopper Nilaparvata lugens is one of the most destructive insect pests in Asia, demonstrating high fertility and causing huge crop losses by sucking sap of rice as well as transmitting viruses. However, functional genomic studies on N. lugens are seriously constrained by lack of genetic tools. Here, we employed two eye pigmentation genes to generate germ-line mutations in N. lugens using the CRISPR/Cas9 (clustered regularly interspaced palindromic repeats/CRISPR-associated) system. We showed that injection of single guide RNA of the cinnabar gene of N. lugens (Nl-cn ) into pre-blastoderm eggs induced insertion and deletion (indels) in the founder generation (G 0), which were heritably transmitted to the following G1 generation, leading to bright red compound eyes and ocelli. Mutations of N. lugens white (Nl-w ) generated a high mutant rate of up to 27.3%, resulting in mosaic eyes consisting of white and lightly pigmented ommatidia in both G 0 and G1 individuals. The specificity of CRISPR/Cas9-mediated mutagenesis was further bolstered by PCR and RNA interference-based knockdown analysis. These results show that CRISPR/Cas9-mediated gene editing is achievable in a hemipteran insect, offering a valuable tool for the study of functional genomics and pest management in this planthopper species.

Central orexinergic system contributes to the regulation of cardiovascular function. Orexinergic neurons receiving projections of nerve fibers from multiple structures of brain which involved in control and regulation of cardiovascular function locate in hypothalamus, and their axon terminals widely project to various central structures where orexins receptors are expressed. Here, we summarize the present knowledge that describes the influence of central orexinergic system on cardiovascular activity, the relevance of dysfunction in central orexinergic system with hypertension and psychological stress induced cardiovascular reactivity which are serious risk factors for cardiovascular disease and cardiovascular death. We propose that central orexinergic system may be potentially important targets for the prevention of cardiovascular disease and cardiovascular death, and different orexinergic system involved neuronal circuits may be involved in distinct cardiovascular functions. Acupuncture having bidirectional regulatory ability and a much lower incidence of side effects can prevent disease. We review the improvement of acupuncture on hypertension and psychological stress induced cardiovascular reactivity. We think that acupuncture intervenes hypertension and psychological stress induced cardiovascular reactivity to prevent cardiovascular disease and cardiovascular death. We also summarize relation between acupuncture and central orexinergic system. We propose a hypothesis that acupuncture improve hypertension and psychological stress induced cardiovascular reactivity through regulating central orexinergic system. The knowledge is beneficial for the development of potential therapeutic targets and methods to prevent cardiovascular disease and cardiovascular death.

Pion generalized parton distributions and light-front wave functions are investigated in the framework of Nambu–Jona-Lasinio model. The properties of pion generalized parton distributions and light-front wave functions are examined separately. Proceeding from generalized parton distributions and light-front wave functions to form factors, parton distribution functions, parton distribution amplitudes and transverse momentum dependent parton distributions, giving a complete picture of pion structure. Moreover, comparing these distributions from the two different methods and examining their peculiarities, finding that the two methods give the same distributions. This means that generalized parton distributions and light-front wave functions give the same multi-dimensional mapping of pion in the Nambu–Jona-Lasinio model. In addition, we studied generalized form factors of pion: the quark pressure distribution θ1 and the quark mass distribution θ2, both are harder than electromagnetic form factor. The tensor anomalous magnetic moment B1,0 and the tensor generalized form factor B2,0 are harder than corresponding lattice data.

The homeotic complex (Hox) gene Ultrabithorax (Ubx) plays pivotal roles in modifying specific morphological differences among the second (T2), the third thoracic (T3), and the first abdomen (A1) segment in several insects. Whether Ubx regulates wing dimorphism and other morphological traits in the delphacid family (order Hemiptera) remains elusive. In this study, we cloned a full-length Ubx ortholog (NlUbx) from the wing-dimorphic planthopper Nilaparvata lugens, and identified two NlUbx isoforms. RNA-interference (RNAi)-mediated silencing of NlUbx in short-winged BPH nymphs significantly induced the development of wing-like appendages from T3 wingbuds, and this effect is likely mediated by the insulin/insulin-like signaling pathway. RNAi knockdown of NlUbx in long-winged BPH nymphs led to a transformation from hindwings to forewings. Additionally, silencing of NlUbx not only dramatically changed the T3 morphology, but also led to jumping defect of T3 legs. First-instar nymphs derived from parental RNAi had an additional leg-like appendages on A1. These results suggest that Ubx plays a role in determining some morphological traits in delphacid planthoppers, and thus help in understanding evolution of morphological characteristics in arthropods.

A new landfill liner system composed of two-layer soils was proposed. The upper soil layer composed of 90% raw clay plus 10% lime contributes significant to bearing capacity of liners, and the lower soil layer containing 3 or 6% granular activated carbon (GAC) or 3 or 6% acid-activated bentonite contributes to adsorption capacity of liners. The results from unconfined compressive strength tests showed that the method of adding lime into liner soil material was effective measure to improve compressive strength of liners. The batch test results indicated that GAC and acid-activated bentonite were two potential materials for use as adsorptive amendments for trapping heavy metal ions in liners. To investigate the transport of Cr(VI) through the new liner system, laboratory column tests were performed. The column tests results indicated that the lower soil layer containing adsorptive amendments prevented effectively the transport of Cr(VI). The diffusion coefficients of Cr(VI) for liner soil materials were back-calculated by a one-dimensional numerical transport program. The diffusion coefficient of Cr(VI) was 4.64 × 10−8 m2/s on average for the upper soil layer. The diffusion coefficient of the lower soil layer was 5.4 × 10−10 m2/s to 7.0 × 10−10 m2/s. The differences of diffusion coefficients may be due to different grain size distributions and moulding density between these soils.

The potential of clays containing 3 or 6% granular activated carbon (GAC) or 3 or 6% acid-activated bentonite was assessed for use as a landfill liner-soil material. Permeability, batch adsorption and column tests were performed to evaluate the seepage properties, the Cr(VI) adsorption capacity and transport parameters of the soils. All soil specimens had a hydraulic conductivity of −7 cm/s at a certain range of moulding water content. The Cr(VI) adsorption was much larger than adsorption by the raw clay. The improved clay prevented effectively the transport of Cr(VI). The diffusion coefficient of Cr(VI) for liner-soil materials was back-calculated by a one-dimensional nonlinear advection–dispersion–reaction numerical transport program. This coefficient for the clay or the improved clay was 2.2 × 10 −10 to 8.3 × 10 −10 m 2 /s. The improved clay may be used as a good barrier material to attenuate contamination of Cr(VI) in landfills.

SummaryCrossveinless 2 (CV-2) is an extracellular BMP modulator protein belonging to the Chordin family. During development it is expressed at sites of high BMP signaling and like Chordin CV-2 can either enhance or inhibit BMP activity. CV-2 binds to BMP-2 via its N-terminal Von Willebrand factor type C (VWC) domain 1. Here we report the structure of the complex between CV-2 VWC1 and BMP-2. The tripartite VWC1 binds BMP-2 only through a short N-terminal segment, called clip, and subdomain (SD) 1. Mutational analysis establishes that the clip segment and SD1 together create high-affinity BMP-2 binding. All four receptor-binding sites of BMP-2 are blocked in the complex, demonstrating that VWC1 acts as competitive inhibitor for all receptor types. In vivo experiments reveal that the BMP-enhancing (pro-BMP) activity of CV-2 is independent of BMP-2 binding by VWC1, showing that pro- and anti-BMP activities are structurally separated in CV-2.

The adsorption capacity of landfill liners containing granular activated carbon (GAC), or bentonite activated by acid, for Cr(VI) was investigated by batch testing. The results show that both GAC and activated bentonite could be used as sorptive amendments for trapping Cr(VI) in landfill liners. The Cr(VI) sorption to GAC and activated bentonite is much greater than Cr(VI) sorption to natural clay. The adsorption capacity of Cr(VI) onto all the soils increases with increasing temperature; adsorption capacity is also significantly influenced by soil-solid concentration. As the soil-solid concentration increases the adsorption capacity first decreases logarithmically, but then stabilizes when the soil-solid concentration exceeds a critical value (e.g. 400 g/L). Permeability tests were conducted in the laboratory. The results indicate that the hydraulic conductivity of landfill liners containing GAC or activated bentonite can meet the engineering requirement of 1 nm/s. One-dimensional transport simulations for Cr(VI) were performed to evaluate the effect of GAC and activated bentonite on landfill liners. The results of the simulations indicate that landfill liners containing GAC, or activated bentonite, significantly retard the transport of Cr(VI) relative to a conventional clay liner.

Bone morphogenetic protein (BMP) function is regulated in the extracellular space by many modulator proteins, including those containing a von Willebrand factor type C (VWC) domain. The function of the VWC domain-containing proteins in development and diseases has been extensively studied. The structural basis, however, for the mechanism by which BMP is regulated by these proteins is still poorly understood. By analyzing chordin, CHL2 (chordin-like 2), and CV2 (crossveinless 2) as well as their individual VWC domains, we show that the VWC domain is a versatile binding module that in its multiple forms and environments can expose a variety of binding specificities. Three of four, two of three, and one of five VWCs from chordin, CHL2, and CV2, respectively, can bind BMPs. Using an array of BMP-2 mutant proteins, it can be demonstrated that the binding-competent VWC domains all use a specific subset of BMP-2 binding determinants that overlap with the binding site for the type II receptors (knuckle epitope) or for the type I receptors (wrist epitope). This explains the competition between modulator proteins and receptors for BMP binding and therefore the inhibition of BMP signaling. A subset of VWC domains from CHL2 binds to the Tsg (twisted gastrulation) protein similar to chordin. A stable ternary complex consisting of BMP-2, CHL2, and Tsg can be formed, thus making CHL2 a more efficient BMP-2 inhibitor. The VWCs of CV2, however, do not interact with Tsg. The present results show that chordin, CHL2, and CV2 regulate BMP-2 signaling by different recognition mechanisms.

Interleukin 21 (IL-21) is a recently identified novel cytokine that plays an important role in the regulation of B, T, and NK cell functions. Its effects depend on binding to and signaling through an IL-21 receptor complex consisting of the IL-21 receptor (IL-21R) and the common gamma-chain (gamma(c)). In this study using biosensor technique, the ligand-binding properties of IL-21R and gamma(c), which are presently poorly understood on a molecular level, were analyzed employing recombinant ectodomains of IL-21R and gamma(c). The formation of a binary complex between IL-21 and immobilized IL-21R (K(D) 70pM), gamma(c) and immobilized IL-21 (K(D) 160 microM) and a ternary complex between gamma(c) and IL-21 saturated immobilized IL-21R (K(D) 160nM) could be analyzed. The gamma(c) residues involved in IL-21 binding were defined by alanine-scanning mutational analysis. The epitope comprises gamma(c) residues N44, Y103, N128, L161, E162, and L208. It is not identical but partially overlapping with the previously established gamma(c) epitope for IL-4 binding. These results open the way to understand the molecular recognition mechanism in the IL-21 receptor system and also the promiscuous binding properties of gamma(c).

Interleukin-4 (IL-4) and IL-13 are the only cytokines known to bind to the receptor chain IL-4Ralpha. Receptor sharing by these two cytokines is the molecular basis for their overlapping biological functions. Both are key factors in the development of allergic hypersensitivity, and they also play a major role in exacerbating allergic and asthmatic symptoms. Knowledge of structure and function of this system has allowed the development of inhibitors that block the interaction between the cytokines and their shared receptor. Mutational analysis of IL-4 has revealed variants with high-affinity binding to IL-4Ralpha but no detectable affinity for the second receptor subunit, which is either (gamma)c or IL-13Ralpha1. These IL-4 antagonists fail to induce signal transduction and block IL-4 and IL-13 effects in vitro. IL-4 antagonists prevent the development of allergic disease in vivo and an antagonistic variant of human IL-4 is now in clinical trials for asthma. Detailed knowledge of the site of interaction of IL-4 and IL-4Ralpha has been gained by structure analysis of the complex of these two proteins and through functional studies employing mutants of IL-4 and its receptor subunits. Based on these new data, the hitherto elusive goal of designing small molecular mimetics may be feasible.