Cartilage-inspired terpolymer hydrogel with excellent mechanical properties and superior lubricating ability.

Osteoarthritis (OA), the most common degenerative joint disorder, seriously affects patients' daily activities. Recently, hydrogels, due to their similar structure to articular cartilage, have shown great potential as cartilage-repairing materials. In the present work, we developed a simple process for fabricating terpolymer [P(acrylamide-co-acrylic acid-co-2-acrylamido-2-methyl-1-propanesulfonic acid)/Fe3+] hydrogel [P(AAm-co-AAc-co-AMPS)/Fe3+]. The content of AMPS was found to show a crucial effect on the mechanical and tribological performance of the terpolymer hydrogel. When the content of AMPS was 0.45 mol L-1, the compressive strength, modulus, and friction coefficient of the terpolymer hydrogel were 66.60 ± 1.79 MPa, 2.10 ± 0.16 MPa, and 0.032, respectively. In addition, the hydrogel showed high wear durability and the friction coefficient was as low as 0.038 after 3.6 × 105 sliding cycles.

Characterisation of a copper/zinc superoxide dismutase from Pieris rapae and its role in protecting against oxidative stress induced by chlorantraniliprole.

Insecticide exposure typically leads to abnormally high levels of reactive oxygen species (ROS) and oxidative damage in insects. Superoxide dismutases (SODs) are potent antioxidant enzymes for ROS scavenging that are essential to protect insects against insecticide-induced oxidative injury. The small white butterfly, Pieris rapae, is an economically important lepidopteran pest of cruciferous crops, and the anthranilic diamide insecticide chlorantraniliprole is widely used to control this organism. However, whether chlorantraniliprole causes oxidative stress, and whether SODs are involved in ROS scavenging, remains unclear in P. rapae. In this study, an intracellular copper/zinc SOD (designated PrSOD1) gene was identified and characterised in P. rapae. The gene consists of four exons and three introns, and the PrSOD1 protein encoded by the gene has typical highly conserved features of CuZnSODs, including two signature motifs and seven Cu/Zn-interacting residues. Transcription of PrSOD1 was highest in the larval fat body and at the fifth-instar larval stage. Recombinant PrSOD1 protein expressed in Escherichia coli displayed antioxidant activity and high thermal and pH stability, confirming that PrSOD1 encodes a functional enzyme. Exposure to three sublethal doses of chlorantraniliprole for 6, 12 or 24 h resulted in significantly increased malondialdehyde concentration in P. rapae larvae, indicating insecticide-induced oxidative stress. Furthermore, both PrSOD1 transcription levels and CuZnSOD activity were quickly (6 and 12 h, respectively) upregulated in larvae subjected to chlorantraniliprole, strongly suggesting that PrSOD1 plays an important role in protecting against oxidative damage and possibly chlorantraniliprole tolerance in P. rapae.

Functional characterization of a binding protein for Type-II sex pheromones in the tea geometrid moth Ectropis obliqua Prout.

The tea geometrid moth Ectropis obliqua Prout is one of the most serious moth pests in tea plants, and its sex pheromones have been identified as typical Type-II polyunsaturated hydrocarbons and epoxide derivatives. Therefore, the E. obliqua male olfactory system provides a good model to study the molecular basis of Type-II sex pheromone recognition as well as functional gene evolution towards structurally different types of moth sex pheromones. In this study, we identified the full-length sequence of a pheromone-binding protein, EoblPBP2 and revealed that it clustered together with the lepidopteran PBP2 subfamily, which binds Type I acetate pheromones. These findings suggest that the EoblPBP2 sequence and physiological function are conserved, although E. obliqua evolved Type II hydrocarbon and epoxide sex pheromones structurally different from Type I acetates. To examine this hypothesis, we studied the expression patterns and in vitro functions of EoblPBP2 in detail. Quantitative real-time PCR experiments showed that EoblPBP2 was predominantly expressed in male E. obliqua antennae. Fluorescence in situ hybridization further demonstrated that the EoblPBP2 gene was abundantly expressed in the pheromone-sensitive sensilla trichodea Str-I in male E. obliqua. The physiological function of recombinant EoblPBP2 was then examined using a competitive binding assay. The results showed that EoblPBP2 had high affinities for three E. obliqua Type II sex pheromone components and Type I acetate pheromones in comparison to some plant volatiles. These results indicate that PBP2 is involved in the detection of Type II pheromones in E. obliqua and it still retains high binding affinities to acetate pheromones and some green leaf ester volatiles.

Molecular characterization of a typical 2-Cys thioredoxin peroxidase from the Asiatic rice borer Chilo suppressalis and its role in oxidative stress.

In insects, thioredoxin peroxidase (TPX) plays an important role in protecting against oxidative damage. However, studies on the molecular characteristics of TPXs in the Asiatic rice borer, Chilo suppressalis, are limited. In this work, a cDNA sequence (CsTpx3) encoding a TPX was identified from C. suppressalis. The deduced CsTPX3 protein shares high sequence identity and two positionally conserved cysteines with orthologs from other insect species, and was classified as a typical 2-Cys TPX. CsTpx3 was expressed most highly during the fifth-instar larval stage, and transcripts were most abundant in the midgut. Recombinant CsTPX3 protein expressed in Escherichia coli displayed the expected peroxidase activity by removing H2 O2 . Furthermore, CsTPX3 protected DNA from oxidative damage, and E. coli cells overexpressing CsTPX3 exhibited long-term resistance to oxidative stress. Exposure to various oxidative stressors, such as cold (8°C), heat (35°C), bacteria (E. coli), and two insecticides (chlorpyrifos and lambda-cyhalothrin), significantly upregulated transcription of CsTpx3. However, exposure to abamectin had no such effect. Our results provide valuable information for future studies on the antioxidant mechanism in this insect species.

Nano Serpentine Powders as Lubricant Additive: Tribological Behaviors and Self-Repairing Performance on Worn Surface.

Natural serpentine powders are applicable as additives for various lubricating oils. However, no uniform theories explain their tribological performance, lubrication, and wear mechanism, especially their self-repairing mechanism. Herein, the influence of different nano serpentine powders (NSPs) contents in liquid paraffin on the friction and wear characteristics of steel balls and the self-repairing process of NSPs on the worn surface were studied. Results show that the optimal amount of NSPs was 0.5 wt %. Relative to those of the base oil, the friction coefficients and wear spot diameters were reduced by 22.8% and 34.2%, respectively. Moreover, the long-term tribological test shows that the wear scar diameter decreased slightly after 3 h, reaching the state of dynamic balance between wear and repair. The outstanding tribological performance should be attributed to the formed bilayer tribofilm, the first layer of which contains nanoparticles surrounded by lubricants and the second layer of which contains nanoparticles compacted onto the surface of the steel ball.

The sensilla trichodea-biased EoblPBP1 binds sex pheromones and green leaf volatiles in Ectropis obliqua Prout, a geometrid moth pest that uses Type-II sex pheromones.

Pheromone-binding proteins (PBPs) are considered to play critical roles in sex pheromone detection. Lepidopteran moths can be divided into two taxa, those that use Type-I sex pheromones, such as C10-C18 unsaturated aldehydes, alcohols and acetates, and those that use Type-II pheromones, which are C17-C23 polyunsaturated hydrocarbons and their epoxide derivatives. To date, nearly all the characterized PBPs have been reported in moths with Type-I sex pheromones, and the physiological functions of PBPs in moths that use Type-II sex pheromones remains unclear. In the present study we functionally examine EoblPBP1 in Ectropis obliqua Prout, an important geometrid moth pest that uses Type-II sex pheromones. The phylogenetic analysis of the sequence indicated that EoblPBP1 clustered together with ScerPBP1, a geometrid PBP for detecting Type-II sex pheromones. Scanning electron microscopy showed that E. obliqua moths of both sexes mainly had six types of antennal sensilla, including two types of sensilla trichodea, Str-I and Str-II, sensilla basiconica (Sba), sensilla styloconica (Sst), sensilla chaetica (Sch) and sensilla auricillica (Sau). Of these, Str-I was confirmed to be male moth-specific and had five different subtypes. Fluorescence in situ hybridization revealed that EoblPBP1 was primarily expressed at the base of Str-I. A comparative binding assay showed that recombinant EoblPBP1 bound three sex pheromone components of E. obliqua, demonstrating its involvement in the detection of Type-II sex pheromones. Besides, EoblPBP1 also highly bound unsaturated acetates pheromones and the green leaf volatiles. These results indicate that PBP1 is associated with detecting Type-II sex pheromones in E. obliqua but cannot differentiate Type-II sex pheromones from Type-I sex pheromones or green leaf volatiles. Our findings provide a foundation for further study on molecular basis of Type-II sex pheromone recognition in lepidopteran moths.

Molecular Characterization of a Mitochondrial Manganese Superoxide Dismutase From Chilo suppressalis (Lepidoptera: Crambidae).

In insects, superoxide dismutases (SODs) play a critical role in the scavenging of harmful reactive oxygen species (ROS) and protecting against oxidative stress induced by various environmental stresses. The Asiatic rice borer, Chilo suppressalis (Walker) (Lepidoptera: Crambidae), is an economically important insect pest of rice crops. In this study, a mitochondrial manganese SOD (Cs-mMnSOD) gene was characterized in C. suppressalis. The deduced Cs-mMnSOD protein has typical highly conserved features of mitochondrial manganese SODs, including four manganese binding residues, the signature DVWEHAYY peptide, and a mitochondrial-targeting sequence at the N-terminus. Transcription of Cs-mMnSOD was detectable at all developmental stages, but highest in pupae. Furthermore, the mRNA level of Cs-mMnSOD was strongly upregulated (more than twofold increase) following exposure to low and high temperatures (4, 30 and 35°C), insecticides (chlorpyrifos and chlorantraniliprole), and chemical reagents (cumene hydroperoxide, paraquat, H2O2 and CdCl2), but slightly elevated (less than twofold increase) in response to 8°C, abamectin and CuSO4. Additionally, the Cs-mMnSOD transcription results were consistent with the enzymatic activity data of the protein product. Purified recombinant Cs-mMnSOD protein expressed in Escherichia coli displayed SOD activity and thermostability. Furthermore, E. coli cells overexpressing Cs-mMnSOD exhibited long-term resistance to the oxidative inducers cumene hydroperoxide and paraquat. Our findings indicate that Cs-mMnSOD plays an important role in protecting C. suppressalis against oxidative damage.

Rapid Biodegradation of the Herbicide 2,4-Dichlorophenoxyacetic Acid by Cupriavidus gilardii T-1.

Phytotoxicity and environmental pollution of residual herbicides have caused much public concern during the past several decades. An indigenous bacterial strain capable of degrading 2,4-dichlorophenoxyacetic acid (2,4-D), designated T-1, was isolated from soybean field soil and identified as Cupriavidus gilardii. Strain T-1 degraded 2,4-D 3.39 times more rapidly than the model strain Cupriavidus necator JMP134. T-1 could also efficiently degrade 2-methyl-4-chlorophenoxyacetic acid (MCPA), MCPA isooctyl ester, and 2-(2,4-dichlorophenoxy)propionic acid (2,4-DP). Suitable conditions for 2,4-D degradation were pH 7.0-9.0, 37-42 °C, and 4.0 mL of inoculums. Degradation of 2,4-D was concentration-dependent. 2,4-D was degraded to 2,4-dichlorophenol (2,4-DCP) by cleavage of the ether bond and then to 3,5-dichlorocatechol (3,5-DCC) via hydroxylation, followed by ortho-cleavage to cis-2-dichlorodiene lactone (CDL). The metabolites 2,4-DCP or 3,5-DCC at 10 mg L-1 were completely degraded within 16 h. Fast degradation of 2,4-D and its analogues highlights the potential for use of C. gilardii T-1 in bioremediation of phenoxyalkanoic acid herbicides.