The Sequence Characteristics and Binding Properties of the Odorant-Binding Protein 2 of Euplatypus parallelus to Semiochemicals.

Euplatypus parallelus is one of the dominant rubber bark beetle species in Hainan's rubber-planting area. Semiochemicals, including the volatiles found in rubber trees and aggregation pheromones, play an important role in the search for suitable host plants. To examine the possible functional role of highly expressed odorant-binding protein 2 of Euplatypus parallelus (EparOBP2) in the semiochemical recognition process, we cloned and analyzed the cDNA sequence of EparOBP2. The results showed that EparOBP2 contains an open reading frame (ORF) of 393 bp that encodes 130 amino acids, including a 21-amino-acid residue signal peptide at the N-terminus. The matured EparOBP2 protein consists of seven α-helices, creating an open binding pocket and three disulfide bridges. The results of the fluorescence binding assay showed that EparOBP2 had high binding ability with α-pinene and myrcene. The docking results confirmed that the interactions of α-pinene and myrcene with EparOBP2 were primarily achieved through hydrophobic interactions. This study provides evidence that EparOBP2 may be involved in the chemoreception of semiochemicals and that it can successfully contribute to the integrated management of E. parallelus.

Immunogenicity of the recombinant adenovirus fusion-expressing E0-E2 gene of the classical swine fever virus.

Adenovirus vector vaccines have been the mainstream research direction of CSF vaccines, due to the replication deficiency of adenovirus vectors, achieving double effects with the safety of inactivated vaccines and the efficacy of live vaccines. Therefore, the E0 and E2 genes were expressed by an adenovirus vector, a recombinant adenovirus E0-E2 (rAd-E0-E2) vaccine was constructed, and the minimum immunization dose and immune duration period were determined in this study. Forty healthy piglets were randomly divided into 8 groups (n = 5). Groups 1 ~ 5 were used to determine the minimum immunization dose, and 5 groups were inoculated with rAd-E0-E2 at different immune doses. Serum was collected at 7 d and 14 d after immunization to detect CSFV antibodies by ELISA, and piglets were challenged at 7 d post immunization. Groups 6 ~ 8 were immunized with 1 dose of rAd-E0-E2, the CSFV live attenuated vaccine C strain and saline to identify the immune duration period. Serum was collected at different time points after immunization, CSFV antibodies were detected by ELISA, and piglets were challenged at 8 months post immunization. Meanwhile, temperature, clinical symptoms and pathology were observed. The results of groups 1 ~ 5 showed that 1 piglet was protected after challenge, and 4 piglets exhibited high fever retention, typical CSFV symptoms and tissue lesions in the 1/50 dose group, whereas no clinical symptoms were observed in the 1/10 dose, 1/5 dose or 1 dose groups with 5/5 protection after challenge. The minimum dose was determined as 1/10 dose. The results of groups 6 ~ 8 showed that all piglets survived after challenge, but the antibody level of the rAd-E0-E2 strain was higher than that of the C strain at 8 months post immunization, and all piglets in the negative group developed the disease process after challenge. Overall, the minimum immunization dose of rAd-E0-E2 was 1/10 dose (3.16 × 106.0 IFU) and the minimum immune dose was determined to be 1 dose (3.16 × 107.0 IFU) to achieve the expected effects. The immune duration period of piglets immunized with 1 dose of rAd-E0-E2 was at least 8 months.

Ultrastructure Characteristics and Sexual Dimorphism of Antennal Sensilla in Tirathaba rufivena (Lepidoptera: Pyralidae).

Tirathaba rufivena Walker, a major insect pest of Areca catechu L., has severely threatened areca nut cultivation in Hainan, China. To improve our understanding of the communication mechanism in host plant seeking and mate-finding for T. rufivena, we described and further characterized the external morphology and internal sensilla structures using scanning electron microscopy and transmission electron microscopy in this study. The antennal morphology was similar between males and females, and there was no significant difference in length between the two sexes. In total, nine sensilla types were identified: sensilla trichodea (Str), sensilla chaetica (Sch), sensilla basiconica (Sba), sensilla auricillica (Sau), sensilla coeloconica (Sco), sensilla styloconica (Sst), Böhm sensilla (Bs), uniporous peg sensilla (Ups) and sensilla squamiformia (Ssq). Sexual dimorphism mainly occurs in variation in the length of Sba, Sch, Sco1 and Bs, and the abundance of Sba, Sau1 and Sau2. The Sba had larger size and numbers on female antennae than that on males, suggesting that these sensilla might have important roles in locating host plants. Both Sau1 and Sau2 were significantly more abundant in females and were probably associated with the detection of mates and host plant for oviposition. These data were important for ongoing studies on host plant seeking and mate-finding behavior in T. rufivena and provided a theoretical foundation to further studies of semiochemical control for this pest.

Binding Properties of Odorant-Binding Protein 4 of Tirathaba rufivena to Areca catechu Volatiles.

Odorant-binding proteins (OBPs) play a key role in the olfactory system and are essential for mating and oviposition host selection. Tirathaba rufivena, a serious lepidopterous insect pest of the palm area in recent years, has threatened cultivations of Areca catechu in Hainan. Female-biased odorant-binding protein 4 of T. rufivena (TrufOBP4) expression was hypothesized to participate in the process of oviposition host recognition and localization. In this study, we cloned and analyzed the cDNA sequence of TrufOBP4. The predicted mature protein TrufOBP4 is a small, soluble, secretory protein and belongs to a classic OBP subfamily. Fluorescence binding assay results showed that TrufOBP4 had high binding abilities with the host plant volatiles, octyl methoxycinnamate, dibutyl phthalate, myristic acid and palmitic acid. These four components tend to dock in the same binding pocket based on the molecular docking result. The interactions and contributions of key amino acid residues were also characterized. This research provides evidence that TrufOBP4 might participate in the chemoreception of volatile compounds from inflorescences of A. catechu and can contribute to the integrated management of T. rufivena.

Magnetic Cell Centrifuge Platform Performance Study with Different Microsieve Pore Geometries.

The detection and analysis of circulating tumor cells (CTCs) plays a crucial role in clinical practice. However, the heterogeneity and rarity of CTCs make their capture and separation from peripheral blood very difficult while maintaining their structural integrity and viability. We previously demonstrated the effectiveness of the Magnetic Cell Centrifuge Platform (MCCP), which combined the magnetic-labeling cell separation mechanism with the size-based method. In this paper, a comparison of the effectiveness of different microsieve pore geometries toward MCCP is demonstrated to improve the yield of the target cell capture. Firstly, models of a trapped cell with rectangular and circular pore geometries are presented to compare the contact force using finite element numerical simulations. The device performance is then evaluated with both constant pressure and constant flow rate experimental conditions. In addition, the efficient isolation of magnetically labeled Hela cells with red fluorescent proteins (target cells) from Hela cells with green fluorescent protein (background cells) is validated. The experimental results show that the circular sieves yield 97% purity of the target cells from the sample with a throughput of up to 2 µL/s and 66-fold sample enrichment. This finding will pave the way for the design of a higher efficient MCCP systems.