Transcriptional response and functional analysis of ATP-binding cassette transporters to tannic acid in pea aphid, Acyrthosiphon pisum (Harris).

Although tannins are widely distributed in broad beans and alfalfa, the pea aphid (Acyrthosiphon pisum) can still destroy them. The ATP binding cassette (ABC) transporters participate in the metabolism of plant secondary metabolites and pesticides in insects. However, whether ABC transporter genes play a role in the metabolism of tannins in the pea aphid is unclear. Here, we found that verapamil (an ABC transporter inhibitor) significantly increased the mortality of tannic acid to pea aphid, which indicated that ABC transporter gene was related to the metabolism of tannic acid by pea aphid. Then, we identified 54 putative ABC transporter genes from the genome database of A. pisum. These genes were divided into eight subfamilies, ApABCA to ApABCH, of which subfamily G has the largest number of genes with 19, followed by the subfamily C with 14. RT-qPCR results show that the expression levels of ApABCA2, ApABCC7, ApABCG2, and ApABCG3 were highly expressed in the first instar, while those of ApABCA3, ApABCG6, ApABCG7, ApABCH3, and ApABCH4 were highly expressed in adults. Furthermore, transcription levels of many ABC transporter genes were induced by tannic acid. Especially, ApABCG17 and ApABCH2 were obviously induced after being exposed to tannic acid. Meanwhile, knockdown of ApABCG17 by RNA interference resulted in increased sensitivity of pea aphid to tannic acid. These results suggest that ApABCG17 may be involved in tannic acid metabolism in pea aphid. This study will help us to understand the mechanism of tannic acid metabolism in pea aphid, and provides a basis for further research on the physiological function of ABC transporter genes in pea aphid.

Citrus fiber for the stabilization of O/W emulsion through combination of Pickering effect and fiber-based network.

In this study, oil-in-water emulsions stabilized by citrus fiber were prepared and characterized. We found that citrus fiber can produce stable gel-like, surfactant-free O/W emulsions with microscale droplet sizes at fiber concentrations upon 2% (W/V) using 25% (V/V) oil. The interfacial framework, citrus fiber partition between the continuous phase and state of the droplets of emulsions were visualized by confocal laser scanning microscopy (CLSM), confirming that in addition to Pickering stabilization, the citrus fiber-based network also contributed to stabilization of the emulsions. The citrus fiber-stabilized emulsion is typical non-Newtonian fluid and its interfacial viscosity is not influenced obviously by changing pH from 2 to 10, ionic strength of NaCl from 0.00 to 1.00 mol/L or temperature from -20 to 70 °C. The acquired findings in this study show that citrus fiber can fabricate Pickering emulsions with excellent stability and solve the problem of resource waste during the pectin produce process.

Comparison of the cervista HPV HR test and luminex XMAP technology for the diagnosis of cervical intraepithelial neoplasia.

OBJECTIVES: This study was designed to compare the Cervista high risk (HR) human papillomavirus (HPV) test with Luminex XMAP technology for the detection of the relationship between HPV infection and cervical intraepithelial neoplasia. METHODS: In total, 3280 patients in a cervical specialty clinic were divided into two groups for either Cervista (1855 patients) or Luminex (1425 patients). Subsequent colposcopy examinations were performed in 1270 women with cytologic results showing ASCUS or higher level lesions. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were evaluated. RESULTS: The positive rates of the Cervista and Luminex groups were 61.48% and 67.43%, respectively, and occurrence in those 30-40 years old was most common. The typing of HPV showed that A9 positive cases were the most prevalent genotype (33.53%), followed by A5/A6 (16.44%) and A7 (11.37%) in the Cervista group, and HPV-16 was the most prevalent genotype (25.81%), followed by HPV-18 (18.6%) and HPV-31 (8.6%) in the Luminex group. Moreover, the overall concordance rate was 96.26% (95% confidence interval, 93.51-99.00%) in the 187 women with cytologic results of ASCUS or higher. There were no significant differences in the positive rates of HPV between the Cervista and Luminex groups, and both had a high sensitivity and NPV. CONCLUSIONS: The results showed a high good concordance between the two methods in diagnostic accuracy. Among the patients in the cervical specialty clinic, both the A9 group of HPV and HPV-16 showed the highest positive rate. Cervista and Luminex shared similar a clinical value in the detection of CIN2 or higher.

[Responses of Arma chinensis cold tolerance to rapid cold hardening and underlying physiological mechanisms].

Rapid cold hardening can enhance the cold tolerance of some insects. To explore the effects of different cold hardening induction temperature on the cold tolerance of Arma chinensis and related physiological mechanisms, the 3rd generation A. chinensis adults reared indoor were treated with cooling at 15, 10, and 4 degrees C for 4 h, respectively, or with gradual cooling from 15 degrees C for 4 h to 10 degrees C for 4 h, and finally to 4 degrees C for 4 h. The super-cooling point, water content, and the contents of low molecular carbohydrates, glycerol, and amino acids of the adults after cooling and the adults cold tolerance at 0, -5, and -10 degrees C were measured by thermocouple, high performance liquid chromatography, and other analytical techniques. When exposed at -10 degrees C after cooling, the survival rate of the adults treated with gradual cooling or treated with cooling at 4 degrees C for 4 h was averagely 58.3%, while that of the adults reared at room temperature (25 degrees C +/- 2 degrees C) or treated with cooling at 15 degrees C or 10 degrees C for 4 h decreased significantly, with an average of 8.9%. The super-cooling point of the adults treated with gradual cooling or with cooling at 4 degrees C for 4 h was -15.6 degrees C, which was averagely 1.3 degrees C lower than that of the other treatments. The water content of the adults had no significant difference among all treatments, with an average of 61.8%, but the glucose, sorbitolum, glycerol, Ala, and Glu contents in treatments gradual cooling and cooling at 4 degrees C for 4 h increased by 2.82-fold, 2.65-fold, 3.49-fold, 51.3%, and 80.2%, while the fucose, mannose, and Pro contents decreased by 68.4%, 52.2%, and 30.2%, respectively, as compared with the other treatments. The fructose content showed no significant difference among all treatments. It was suggested that rapid cool hardening had a critical temperature to induce the physiological metabolism process of adult A. chinensis, and gradual cooling hardening could not further increase the cold tolerance of adult A. chinensis on the basis of rapid cool hardening.

Aggregation pheromone of the Qinghai spruce bark beetle, Ips nitidus eggers.

Volatiles from hindgut extracts of males of the Qinghai spruce bark beetle, Ips nitidus, from different attack phases (phase 1: unpaired males and phases 2-4: males joined with one to three females) and hindgut extracts of mated females were analyzed by gas chromatography-mass spectrometry (GC-MS)/flame ionization detection (FID) with both polar and enantioselective columns. The GC-MS/FID analyses demonstrated that unpaired males from attack phase 1 (nuptial chamber constructed) produced 2-methyl-3-buten-2-ol, approx. 74%-(-)-ipsdienol, and (-)-cis-verbenol as major hindgut components, and (-)-trans-verbenol, (-)-ipsenol, (-)-verbenone, myrtenol, and 2-phenylethanol as minor or trace components. The quantities of 2-methyl-3-buten-2-ol and especially ipsdienol decreased after mating during phases 2-4, whereas the quantities of (-)-cis- and (-)-trans-verbenol did not change. In contrast, the quantity of (-)-ipsenol seemed to increase as mating activity progressed. After mating with three females (harem size = 3; phase 4), only trace to small amounts of male-specific compounds were detected from I. nitidus male hindguts. Chemical analysis of the hindgut extracts of mated females showed only trace amounts of semiochemicals. A field-trapping bioassay in Qinghai, China showed that the four-component "full blend" containing the three major components, 2-methyl-3-buten-2-ol, (+/-)-ipsdienol, and (-)-cis-verbenol, plus a minor component, (-)-trans-verbenol, caught significantly more I. nitidus (male/female = 1:2.2) than did the unbaited control and two binary blends. The replacement of (+/-)-ipsdienol with nearly enantiomerically pure (-)-ipsdienol in the "full blend" significantly reduced trap catches, which suggests that both enantiomers are needed for attraction. On the other hand, removal of (-)-trans-verbenol from the active "full blend" had no significant effect on trap catches. Our results suggest that the three major components, 2-methyl-3-buten-2-ol, 74%-(-)-ipsdienol, and (-)-cis-verbenol (at 7:2:1), produced by unpaired fed males, are likely the aggregation pheromone components of I. nitidus, thus representing the first characterization of an aggregation pheromone system of a bark beetle native solely to China.

[Immunogenicity of recombinant human zona pellucida-3 peptides expressed in E. coli and efficacy of their antisera to inhibit in vitro human sperm-egg binding].

The present study was aimed to analyze the immunogenicity of recombinant human zona pellucida-3 peptides (r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348)) expressed in E. coli through immunizing rabbits, and to evaluate the efficacy of their polyclonal antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding respectively. Male New Zealand rabbits were immunized using r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) as antigen respectively, which was purified through an improved method of preparative gel polyacryulamide gel electrophoresis. The antibody response level of r-huZP3a(22 approximately 176) or r-huZP3b(177 approximately 348) immunogen in rabbits was determined by ELISA using mouse ZP3-5 (amino acid sequence(137 approximately 150) being completely conserved with huZP3(138 approximately 151) sequence) and specific huZP3-14 (amino acid sequence(327 approximately 340)) synthetic peptides as coating antigens respectively. The immunoreactivity and specificity of the anti-r-huZP3a(22 approximately 176) and anti-r-huZP3b(177 approximately 348) antisera with each r-huZP3 peptides, were tested by immunoblot and immunohistochemistry (using native huZP and human ovary section) respectively. A competitive hemizona assay (HZA) was used to evaluate the efficacy of the antisera against r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) to inhibit in vitro human sperm-egg binding. Both r-huZP3 peptides were able to induce higher antibody titers in rabbits. Each antiserum could specifically recognize or bind to each target r-huZP3 peptide expressed in E. coli and native human ZP in vitro. The antisera also inhibited sperm-egg binding in the HZA. These results show that r-huZP3a(22 approximately 176) and r-huZP3b(177 approximately 348) are of strong immunogenicity. They can be used to develop a kit for detecting whether there are autoantibodies to zona pellucida in unexplained infertile women, and their antisera might be useful tools for determining minimal B-cell epitope sequences of several known huZP3 epitope peptides.

[Advances in molecular mechanisms of human sperm-egg interaction and infertility].

Human sperm-egg recognition, adhesion and fusion are key steps in the whole reproductive process. Some abnormalities in human gamete interaction have been shown to be due to defects in the sperm, others attributed to defects in the zona pellucida (ZP) and the egg plasma membrane. This paper reviews the molecular basis and molecular mechanisms of human sperm-egg interaction. More and more advances in the studies of these aspects are shown to be of significant value to the diagnosis and treatment of infertility as well as to the development of assisted reproductive techniques.