Identification and Structure of Epitopes on Cashew Allergens Ana o 2 and Ana o 3 Using Phage Display.

BACKGROUND: Cashew (Anacardium occidentale L.) is a commercially important plant. Cashew nuts are a popular food source that belong to the tree nut family. Tree nuts are one of the eight major food allergens identified by the Food and Drug Administration in the USA. Allergies to cashew nuts cause severe and systemic immune reactions. Tree nut allergies are frequently fatal and are becoming more common. AIM: We aimed to identify the key allergenic epitopes of cashew nut proteins by correlating the phage display epitope prediction results with bioinformatics analysis. DESIGN: We predicted and experimentally confirmed cashew nut allergen antigenic peptides, which we named Ana o 2 (cupin superfamily) and Ana o 3 (prolamin superfamily). The Ana o 2 and Ana o 3 epitopes were predicted using DNAstar and PyMoL (incorporated in the Swiss-model package). The predicted weak and strong epitopes were synthesized as peptides. The related phage library was built. The peptides were also tested using phage display technology. The expressed antigens were tested and confirmed using microtiter plates coated with pooled human sera from patients with cashew nut allergies or healthy controls. RESULTS: The Ana o 2 epitopes were represented by four linear peptides, with the epitopes corresponding to amino acids 108-111, 113-119, 181-186, and 218-224. Furthermore, the identified Ana o 3 epitopes corresponding to amino acids 10-24, 13-27, 39-49, 66-70, 101-106, 107-114, and 115-122 were also screened out and chosen as the key allergenic epitopes. DISCUSSION: The Ana o 3 epitopes accounted for more than 40% of the total amino acid sequence of the protein; thus, Ana o 3 is potentially more allergenic than Ana o 2. CONCLUSIONS: The bioinformatic epitope prediction produced subpar results in this study. Furthermore, the phage display method was extremely effective in identifying the allergenic epitopes of cashew nut proteins. The key allergenic epitopes were chosen, providing important information for the study of cashew nut allergens.

The influencing factors of Harbin (China) residents' satisfaction with municipal solid waste treatment.

China is experiencing an enormous increase in municipal household solid waste (MHSW) generation and is facing multiple problems associated with the treatment of MHSW. This paper analyses factors affecting residents' satisfaction with MHSW treatment performance. Six factors were identified by the Delphi method: (a) pick-up frequency by waste collection vehicles, (b) fund supply situation, (c) charging standard for waste treatment, (d) waste bin arrangement, (e) laws and regulations, (f) publicity and education. We examine the significance of these six influencing factors, estimating binary logistic regression models. Data for this study are derived from the survey responses of 469 households in Harbin, one of the largest cities in northeast China. The results indicate that 'pick-up frequency by waste collection vehicles' is ranked the first and most important determinant of Harbin residents' satisfaction with MHSW treatment; this is closely followed by 'publicity and education'. The third and fourth significant influencing factors, respectively, are 'fund supply situation' and 'charging standard for waste treatment'. The last two factors are 'laws and regulations' and 'waste bin arrangement'. By understanding the influence of various factors on residents' satisfaction, this study aims to help in designing an effective waste management system to reduce the cost of MHSW management, and to raise the residents' satisfaction with municipal solid waste treatment. Based on the research findings, we advocate that establishing a reasonable waste transport (pick-up) system as well as strengthening publicity and education of waste management are key to improving residents' satisfaction with the MHSW treatment performance.

Deletion of SMURF 1 represses ovarian cancer invasion and EMT by modulating the DAB2IP/AKT/Skp2 feedback loop.

SMAD ubiquitination regulatory factor 1 (SMURF1) has been described as a tumor suppressor in multiple aggressive cancers. Nevertheless, the potential role of SMURF1 in ovarian cancer invasion and epithelial-to-mesenchymal transition (EMT) remains unclear. The aim of this study was to evaluate the efficacy of SMURF1 on tumor migration and EMT and elucidate the underlying molecular mechanism in ovarian carcinoma. We found elevated SMURF1 in several ovarian cancer cells in both messenger RNA and protein. Additionally, silencing SMURF1 apparently repressed cell proliferation and invasion capacity of SKOV3 and A2780 cells and markedly attenuated expression of linked proteins such as proliferating cellnuclear antigen, matrix metalloproteinase (MMP)-2, and MMP-9. Furthermore, depletion of SMURF1 dramatically impeded EMT progress by modulating EMT biomarkers, with a notable increase in E-cadherin expression accompanied by the decrease in N-cadherin and vimentin in both SKOV3 and A2780 cells. Interestingly, elimination of SMURF1 led to disabled homolog 2 DOC-2/DAB2 interacting protein (DAB2IP) activation and dampened AKT/Skp2 signaling. Most important, depleted of DAB2IP or treatment with the AKT agonist 740Y-P effectively abolished the suppressive effects of SMURF1 knockout on cell invasiveness and EMT process. Taken all data together, these findings demonstrated that the absence of SMURF1 repressed cell proliferation, invasive capability, and EMT process in ovarian cancer through DAB2IP/AKT/Skp2 signaling loops, suggesting that SMURF1 may serve as a new potential therapeutic agent for ovarian cancer.

[Production and mechanism of CCL5 by macrophages in U14 cervical cancer-bearing mice during infection].

OBJECTIVE: To investigate the production and mechanism of chemokine (C-C motif) ligand 5 (CCL5) by macrophages in U14 cervical cancer-bearing mice during infection. METHODS: The U14 cervical cancer cells were injected in C57BL/6 mice to induce tumor-bearing condition. Lipopolysaccharide (LPS) was injected into C57BL/6 mice to induce infection. The protein expression of CCL5 in the serum and the CCL5 mRNA expression in inflammatory cells were measured by ELISA and fluorescence quantitative-PCR in four groups. Macrophages were induced in the tumor conditioned medium (TCM) which extracted from mice serum. The protein expression levels of CCL5, prostaglandin E2 (PGE2) and cyclic adenosine monophosphate (cAMP) in the medium and CCL5, PGE2 and cAMP mRNA expression in the macrophages were detected in different groups. In order to determine whether the inhibition was related to PGE2, selective cyclooxygenase 2(COX-2) inhibitor NS398 was used to reverse this phenomenon and protein kinase A (PKA) inhibitor H89 demonstrated the mechanism through blocking cAMP/PKA signaling pathway. RESULTS: (1) The protein and mRNA level of CCL5 in tumor-bearing mice were respectively (151 ± 35) pg/ml and 1.0, which were lower than those in the tumor-free mice (691 ± 85) pg/ml and 4.5 ± 0.8, there were significant difference between them (all P < 0.05). The protein and mRNA level of PGE2 in tumor-bearing mice were (1 198 ± 83) pg/ml and 5.8 ± 0.8, which were higher than those in the tumor-free mice (187 ± 25) pg/ml and 1.0, the difference were significant (all P < 0.05). The protein and mRNA level of CCL5 in tumor-free + LPS mice were (4 049 ± 141) pg/ml and 31.5 ± 2.0, which were higher than those in the tumor-bearing + LPS mice (1 951 ± 71) pg/ml and 12.1 ± 2.8, the difference were also significant (P < 0.05). The protein and mRNA level of PGE2 in tumor-free + LPS mice were (676 ± 70) pg/ml and 3.4 ± 0.4, which were lower than those in tumor-bearing + LPS mice (2 550 ± 382) pg/ml and 11.6 ± 0.9, the difference were also significant (all P < 0.05). (2) Macrophages were cultured in vitro using TCM derived from mice. The protein and mRNA level of CCL5 in tumor-bearing mice TCM were respectively (1 626 ± 177) pg/ml and 28.6 ± 1.2, which were higher than those in the tumor-free mice TCM [(27 ± 3) pg/ml and 1.0], there were significant difference (P < 0.05). The protein and mRNA level of PGE2 in tumor-bearing mice TCM were (790 ± 156) pg/ml and 1.7 ± 0.3, which were higher than those in the tumor-free mice TCM [(448 ± 115) pg/ml, 1.0], the difference were significant (all P < 0.05). The protein and mRNA level of cAMP in tumor-bearing mice TCM were (164 ± 30) pg/ml and 1.6 ± 0.3, which weres higher than those in the tumor-free mice TCM [(118 ± 25) pg/ml,1.0], the difference were significant (all P < 0.05). The protein and mRNA level of CCL5 in tumor-free + LPS mice TCM were (10 475 ± 742) pg/ml and 212.0 ± 5.7, which were higher than those in the tumor-bearing + LPS mice TCM [(6 375 ± 530) pg/ml, 142.3 ± 2.5], the difference were significant (all P < 0.05). The protein and mRNA level of PGE2 in tumor-free + LPS mice TCM were (2 438 ± 95) pg/ml and 4.3 ± 0.7, which weres lower than those in the tumor-bearing + LPS mice TCM [(3 441 ± 163) pg/ml, 5.9 ± 0.3], the difference were significant (all P < 0.05). The protein and mRNA level of cAMP in tumor-free + LPS mice TCM were (340 ± 13) pg/ml and 4.1 ± 0.4, which were lower than those in the tumor-bearing + LPS mice TCM [(542 ± 42) pg/ml, 5.4 ± 0.5], the difference were significant (all P < 0.05). (3) Using COX-2 inhibitor NS398 in the tumor-bearing + LPS mice, the protein and mRNA level of CCL5, PGE2 and cAMP were (7 691 ± 269) pg/ml and 159.0 ± 8.9, (2 820 ± 152) pg/ml and 4.9 ± 0.3, (465 ± 8) pg/ml and 4.3 ± 0.4, respectively, and there were significant difference (all P < 0.05), compared to before treatment. Using PKA inhibitor H89 in the tumor-bearing + LPS mice, the protein and mRNA level of CCL5, PGE2 and cAMP were (8 375 ± 520) pg/ml and 177.0 ± 8.8, (2 650 ± 35) pg/ml and 4.7 ± 0.4, (368 ± 13) pg/ml and 3.1 ± 0.7, respectively, and there were significant difference (all P < 0.05), compared to before treatment. CONCLUSION: TCM of U14 cells activated macrophages to release PGE2 could inhibit the expression of CCL5 levels by cAMP/PKA signaling pathway.

A stochastic frontier analysis of the efficiency of municipal solid waste collection services in China.

Governments in many developing countries, such as China, are investing a lot of human, financial and material resources to ensure that as much of municipal solid waste (MSW) as possible is collected for centralized harmless treatment and disposal. This is regarded as an effective way to alleviate the "waste siege" problem in governance caused by the continuous enormous increase in MSW quantity. Therefore, how to improve the efficiency of MSW collection service system is an important governance issue because the inputs that can be utilized to manage MSW problem are limited due to budget and resource constraints. However, studies on MSW collection efficiency in developing countries like China are under-represented in the existing literature. This paper applies a stochastic frontier analysis (SFA) approach to evaluate the efficiency of MSW collection services in 30 provinces of China from 2008 to 2017, and explore the relative importance of several factors that might influence collection efficiency. The results indicate that there exists considerable room (62.8%) to enhance efficiency since its current efficiency value is merely 0.372. With regard to influencing factors, the proportion of population aged 15-64 appears to have the greatest positive impact on efficiency, along with per capita GDP, added value of tertiary industry and education level. This paper also explores spatial variations of MSW collection efficiency across the eastern, central and western regions. These findings have policy implications and can inform the related government departments how to formulate proper policies to improve collection efficiency.

[Effects of traditional Chinese drugs Shuganyishen, on Ishikawa cell line].

OBJECTIVE: To study the effects of the traditional Chinese drugs, Shuganyishen, on Ishikawa cell line. METHOD: Rat blood serum was prepared by using Chinese drugs serum pharmacology method, in which, Shuganyishen was contained or not contained. Ishikawa cells were subjected into five groups: (1) normal cells group (NCG), (2) rat blood serum without Shuganyishen group (NSGG) as negative control, (3) Shuganyishen group (SGG), (4) Cisplatin group (DDPG) as positive control, and (5) Shuganyishen with Cisplatin group (S&CG). MTT assay was used to evaluate the inhibition effect on cell growth in each group. The mRNA expression of estrogen receptor alpha and beta (ERalpha and ERbeta) in each group was detected by RT-PCR. RESULT: Effective inhibition of cell growth was found in the groups of SGG and S&CG, respectively (P < 0.05), which had no significant difference from positive control by MTT. The mRNA expression of ERbeta was slightly going up, while the expression of ERalpha was hardly effected in SGG by RT-PCR. Interestingly in S&CG, the mRNA expression of ERalpha was significantly down-regulated (P < 0.05), but the mRNA expression of ERbeta remained no change. As the positive control, the mRNA expression of ERalpha and ERbeta was significantly down-regulated in DDPG (P < 0.5). CONCLUSION: Traditional Chinese drugs, Shuganyishen, may inhibit Ishikawa cell growth and had no effects on the expression of ERalpha, furthermore, can reverse the inhibition of ERbeta expression by Cisplatin.

Quantitative evaluation of heavy metals' pollution hazards and estimation of heavy metals' environmental costs in leachate during food waste composting.

Heavy metals in leachate during food waste composting may produce different degrees of pollution hazards and further induce environment costs, when the concentrations of heavy metals exceed the discharging quality standards. Quantitative evaluation of heavy metals' pollution hazards and estimation of such environmental costs are under-represented in the existing literature. This paper uses a logistic function approach to evaluate the extent of pollution hazards of heavy metals such as cadmium (Cd), chromium (Cr), mercury (Hg), lead (Pb) and arsenic (As) and to estimate heavy metals' environmental costs in leachate during food waste composting from Minhang food waste treatment plant located in northern Shanghai, China. Major findings of this study are: (1) The pollution hazards rate of Cd amounts to 94.03%, probably because Cd-containing materials such as plastics are mixed with food waste; (2) With the comprehensive pollution hazards rate estimated as 94.48%, the environmental costs caused by heavy metals in leachate during food waste composting amount to US$0.52 per tonne. This magnitude of environmental costs is meaningful and significant, considering that it is equivalent to 2.97% of Shanghai's food waste treatment charges.

Dandelion-like Co3O4 mesoporous nanostructures supported by a Cu foam for efficient oxygen evolution and lithium storage.

Novel dandelion-like Co3O4 mesoporous nanostructures, supported by a Cu foam, are prepared by a combination of hydrothermal synthesis and annealing. The resulting Co3O4@Cu foam exhibits superior oxygen evolution (Tafel slope = 42.8 mV dec-1) and lithium storage (capacity = 882 mA h g-1@2C after 100 cycles, 1C = 890 mA g-1) properties, which highlight its great promise in the fields of energy storage and conversion.

How beta diversity and the underlying causes vary with sampling scales in the Changbai mountain forests.

This study aims to establish a relationship between the sampling scale and tree species beta diversity temperate forests and to identify the underlying causes of beta diversity at different sampling scales. The data were obtained from three large observational study areas in the Changbai mountain region in northeastern China. All trees with a dbh ≥1 cm were stem-mapped and measured. The beta diversity was calculated for four different grain sizes, and the associated variances were partitioned into components explained by environmental and spatial variables to determine the contributions of environmental filtering and dispersal limitation to beta diversity. The results showed that both beta diversity and the causes of beta diversity were dependent on the sampling scale. Beta diversity decreased with increasing scales. The best-explained beta diversity variation was up to about 60% which was discovered in the secondary conifer and broad-leaved mixed forest (CBF) study area at the 40 × 40 m scale. The variation partitioning result indicated that environmental filtering showed greater effects at bigger grain sizes, while dispersal limitation was found to be more important at smaller grain sizes. What is more, the result showed an increasing explanatory ability of environmental effects with increasing sampling grains but no clearly trend of spatial effects. The study emphasized that the underlying causes of beta diversity variation may be quite different within the same region depending on varying sampling scales. Therefore, scale effects should be taken into account in future studies on beta diversity, which is critical in identifying different relative importance of spatial and environmental drivers on species composition variation.

Lactobacilli inhibit cervical cancer cell migration in vitro and reduce tumor burden in vivo through upregulation of E-cadherin.

The present study was designed to investigate the antitumor effects of Lactobacillus and the potential mechanisms. Cell Counting Kit-8 (CCK-8) assays were carried out to determine suitable doses for investigating the inhibitory effect of lactobacilli on cell migration ability of HeLa and U14 cells in vitro. In addition, western blot assays were performed to investigate the possible mechanisms corresponding to its antitumor effects. Furthermore, a xenograft mouse model was established for investigating the E-cadherin expression in tumor tissues after treatment with lactobacilli. Our results showed that live lactobacilli [multiplicity of infection (MOI) of 1,000:1] significantly possessed inhibitory effects on cell migration ability of cervical cancer cells. Lactobacilli (MOI: 1,000:1) significantly upregulated E-cadherin expressions in HeLa and U14 cells (p<0.05). On the contrary, our results showed that inactivated lactobacilli could not affect the E-cadherin expression levels in HeLa and U14 cells. Similar to the western blot assay, immunohistochemistry results also indicated that lactobacilli treatment significantly upregulated E-cadherin in tumor tissues (p<0.05). In conclusion, our results above suggest that lactobacilli have the potential for inhibiting the migratory ability of cervical cancer cell lines, and the possible pharmacological mechanism may be closely related to the upregulation of E-cadherin.

[Effects of light intensity heterogeneity in gaps of broadleaved Korean pine forest in Changbai Mountains on Pinus koraiensis seedings growth].

By using a Li-6400 portable photosynthesis system, this paper studied the heterogeneity of light intensity in four different size gaps of a broadleaved Korean pine forest in Changbai Mountains, and analyzed the diurnal change of the photosynthesis of Pinus koraiensis saplings in the gaps. In the nine orientations within the gaps, the peak value of the photosynthetically active radiation (PAR) varied in the sequence of west of actual gap, north of extended gap, gap center > south of actual gap, south of extended gap, east of extended gap, east of actual gap > west of extended gap, north of actual gap. Light distribution was dissymmetry in the orientations of east-west and south-north, with the variation in west and north being more significant than that in other orientations. There was no significant difference in the average PAR among the positions within specific orientations. The average PAR of the four gaps from I to IV was 21.85, 45.57, 66.02, and 23.48 micromol x m(-2) x s(-1), respectively, and the difference was statistically significant (P < 0.05). PAR had a significant positive correlation with net photosynthetic rate (P(n)), and the correlation coefficient increased with increasing PAR. With the increase of gap size, both the PAR and the P(n) of P. koraiensis saplings increased first and decreased then, with the maximum values appeared at 267 m2 of gap size.