A novel immunoproteomics method for identifying in vivo-induced Campylobacter jejuni antigens using pre-adsorbed sera from infected patients

Background

Campylobacter jejuni is an important food-borne and zoonotic pathogen with a worldwide distribution. Humans and chickens are hosts of this pathogen. At present, there is no ideal vaccine for controlling human campylobacteriosis or the carriage of C. jejuni by chickens. Bacterial in vivo-induced antigens are useful as potential vaccine candidates and biomarkers of virulence.

Methods

In this study, we developed a novel systematic immunoproteomics approach to identify in vivo-induced antigens among the total cell proteins of C. jejuni using pre-adsorbed sera from patients infected with C. jejuni.

Results

Overall, 14 immunoreactive spots were probed on a PVDF membrane using pre-adsorbed human sera against C. jejuni. Then, we excised these protein spots from a duplicate gel and identified using MALDI–TOF MS. In total, 14 in vivo-induced antigens were identified using PMF and BLAST analysis. The identified proteins include CadF (CadF-1 and CadF-2), CheW, TufB, DnaK, MetK, LpxB, HslU, DmsA, PorA, ProS, CJBH_0976, CSU_0396 and hypothetical protein cje135_05017. Real-time RT-PCR was performed on 9 genes to compare their expression levels in vivo and in vitro. The data showed that 8 of the 9 analyzed genes were significantly upregulated in vivo relative to in vitro.

Conclusion

We successfully developed a novel immunoproteomics method for identifying in vivo-induced Campylobacter jejuni antigens by using pre-adsorbed sera from infected patients.

General significance

This new analysis method may prove to be useful for identifying in vivo-induced antigens within any host infected by bacteria and will contribute to the development of new subunit vaccines.     

The Effect of a LYSE Exporter Overexpression on l-Arginine Production in Corynebacterium crenatum

A targeted approach for direct topical antimicrobial delivery involving the formulation of impregnated freeze-dried wafers prepared from a natural polymer has been assessed to consider potential for treatment of wounded skin. The synthetic cationic antimicrobial peptides (CAPs) NP101 and NP108 were found to have modest in vitro activity against bacterial species commonly associated with wound infections. Minimum inhibitory concentration/minimum bactericidal concentrations against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa were found to be 0.31 mg/ml for NP101 and 0.25–0.5 mg/ml for NP108. Rapid, substantial cytoplasmic potassium loss was induced by NP108 in E. coli, but not the other species. Through scanning electron microscopy, both CAPs were observed to alter cell morphology, prevent normal septation, promote cell aggregation and trigger release or formation of extracellular filaments. Wafers harbouring these agents displayed substantial antibacterial activity when assessed by standard diffusion assay. These data confirm that topical delivery of CAPs, through their incorporation within freeze-dried wafer formulations prepared from natural polymers, represents a potential viable approach for treating skin infection.     

Association between -251A>T polymorphism in the interleukin-8 gene and oral cancer risk: A meta-analysis

Background

Emerging evidence showed that the most common functional polymorphism (-251A>T, rs4073) in the promoter region of the interleukin-8 (IL-8) gene is involved in the regulation of the activities of interleukin-8, thus increasing an individual's susceptibility to oral cancer; but individually published results are inconclusive. The aim of this meta-analysis was to investigate the associations between IL-8 -251A>T polymorphism and oral cancer risk.

Methods

The PubMed, Embase, Web of Science and CBM databases were searched for all articles published up to October 1st, 2012 that addressed IL-8 -251A>T polymorphism and oral cancer risk. Statistical analyses were performed using STATA 12.0 software. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strength of associations.

Results

Six case–control studies were included with a total of 1324 oral cancer cases and 1879 healthy controls. When all available studies were pooled into the meta-analysis, the results showed that the AA and AT genotypes of IL-8 -251A>T polymorphism were associated with increased risk of oral cancer (OR = 1.23, 95% CI: 1.03–1.46, P = 0.025; OR = 1.25, 95% CI: 1.07–1.47, P = 0.006; respectively). In the subgroup analysis by ethnicity, significant associations were observed between the AA and AT genotypes of IL-8 -251A>T polymorphism and increased risk of oral cancer among Caucasian populations (OR = 1.40, 95% CI: 1.14–1.72, P = 0.001; OR = 1.29, 95% CI: 1.06–1.57, P = 0.011; respectively). However, no statistically significant associations were found between IL-8 -251A>T polymorphism and oral cancer risk among Asian populations.

Conclusions

Results from the current meta-analysis indicate that the AA and AT genotypes of IL-8 -251A>T polymorphism might increase the risk of oral cancer, especially among Caucasian populations.     

莫莫格自然保护区白鹤秋季迁徙停歇期觅食生境选择

2008年秋季(10月8日—20日)及2009年秋季(9月20日—10月16日),通过样方法对觅食生境11个生态因子进行调查,利用卡方检验、资源选择指数和资源选择函数在莫莫格保护区对秋季迁徙停歇期白鹤觅食生境选择进行研究。结果表明,白鹤对距人为干扰源距离、植被密度、盖度、高度、植物性食物密度以及水深均具有选择性,但对宏观尺度干扰因子的选择性较低。其偏好觅食生境的特点为:距一级路>5000m,>二级路1500m以上,>三级路1000m以上,>居民点1000m以上,农田>1000m;植被密度20~50株/m2,盖度<10%,高度<20cm,扁杆藨草密度1~50株/m2,藨草密度1~10株/m2,水深40~60cm。白鹤秋季觅食生境资源选择函数为Logistic(P)=0.663+0.565×与一级道路距离+0.042×与二级道路距离+0.519×与三级道路距离+0.353×与居民点距离+0.169×与农田距离–0.455×植被密度–0.618×植被盖度–0.548×植被高度–0.158×扁杆藨草密度–0.404×藨草密度+0.920×水深,T(x)=eLogistic(p)/[1+eLogistic(p)],模型正确预测率为82.9%。     

以葡萄糖为底物一步法合成γ-氨基丁酸整合型重组钝齿棒杆菌的构建

[目的]为了构建一株直接利用廉价的葡萄糖合成γ-氨基丁酸的重组钝齿棒杆菌,将来自于植物乳杆菌γ-氨基丁酸合成途径的关键酶谷氨酸脱羧酶基因(lpgad)在产谷氨酸菌株钝齿棒杆菌中进行整合表达,实现葡萄糖到GABA的一步法生产.[方法]运用PCR技术扩增得到带有tac启动子的谷氨酸脱羧酶基因tacgad.通过重叠PCR的方法获得钝齿棒杆菌精氨酸合成途径关键酶N-乙酰谷氨酸激酶(NAGK)基因内部缺失型基因△argB.利用自杀载体pK18mobsacB构建同源整合载体pK18-△argB::tacgad,以△argB的上下游序列为同源臂,通过两次同源重组将tacgad基因整合到钝齿棒杆菌基因组,同时将NAGK基因argB灭活,利用蔗糖致死基因sacB反向筛选标记筛选得到谷氨酸脱羧酶的重组钝齿棒杆菌C.crenatum △argB::tacgad.重组钝齿棒杆菌以葡萄糖为底物进行发酵,测定GABA含量.[结果]重组菌C.crenatum △argB::tacgad成功表达谷氨酸脱羧酶,同时阻断了精氨酸合成途径对谷氨酸到GABA代谢途径的竞争,粗酶液基本检测不到NAGK活性,发酵液无精氨酸合成.通过96 h发酵,重组菌可积累约8.28 g/L的GABA.[结论]本研究通过将谷氨酸脱羧酶基因定向整合到钝齿棒杆菌精氨酸合成途径的关键酶基因argB内部,成功表达谷氨酸脱羧酶的同时阻断竞争途径精氨酸的合成.本研究为实现直接利用葡萄糖合成GABA的一步法生产奠定了基础.     

Combined small RNA and degradome sequencing reveals microRNA regulation during immature maize embryo dedifferentiation

Genetic transformation of maize is highly dependent on the development of embryonic calli from the dedifferentiated immature embryo. To better understand the regulatory mechanism of immature embryo dedifferentiation, we generated four small RNA and degradome libraries from samples representing the major stages of dedifferentiation. More than 186 million raw reads of small RNA and degradome sequence data were generated. We detected 102 known miRNAs belonging to 23 miRNA families. In total, we identified 51, 70 and 63 differentially expressed miRNAs (DEMs) in the stage I, II, III samples, respectively, compared to the control. However, only 6 miRNAs were continually up-regulated by more than fivefold throughout the process of dedifferentiation. A total of 87 genes were identified as the targets of 21 DEM families. This group of targets was enriched in members of four significant pathways including plant hormone signal transduction, antigen processing and presentation, ECM-receptor interaction, and alpha-linolenic acid metabolism. The hormone signal transduction pathway appeared to be particularly significant, involving 21 of the targets. While the targets of the most significant DEMs have been proved to play essential roles in cell dedifferentiation. Our results provide important information regarding the regulatory networks that control immature embryo dedifferentiation in maize.     

基于MAXENT模型的贺兰山岩羊生境适宜性评价

生境评价对物种保护具有非常重要的意义。本研究在GIS空间技术的支持下,于2010年到2011年间,在贺兰山岩羊分布区采集其地理分布点数据以及环境变量数据,并运用MAXENT模型对贺兰山区域岩羊的生境适宜性进行评价分析。模型的评价结果达到优秀水平,研究结果表明：岩羊生境选择的主要影响因子为矿区、坡度、海拔和道路,生境适宜区主要分布于贺兰山东坡（宁夏贺兰山国家级自然保护区）的西南部,而西坡（内蒙古贺兰山国家级自然保护区）有少量分布,适宜生境面积总和为1 006.9 km2,占研究区域面积的28%。岩羊偏爱于海拔1 500－2 300 m的山势陡峭地带,建议对矿区和道路这两种人为干扰因素采取相应的措施,以提高岩羊的生境质量,促进其种群发展。     

干旱荒漠区银白杨树干液流动态

应用热扩散式树干茎流计(TDP)于2012年7月1日至7月25日,在克拉玛依地区农业开发区对银白杨(Populus alba L.×P.talassica)人工林树干液流速率进行了连续测定,并对气象、土壤水分等指标进行了同步测定。结果表明:7月份的晴天银白杨树干液流速率日变化呈单峰型,阴天呈多峰型,在测量时期液流速率日平均值为0.6059 L/h;银白杨树干单位边材面积的液流速率与太阳总辐射、大气温度、水汽压差呈极显著正相关关系,与相对湿度呈负相关关系。其相关系数绝对值顺序为太阳总辐射>大气温度>水汽压差>相对湿度>风速;银白杨边材面积与胸径之间存在着显著的线性相关关系,相关系数为0.834,单位边材面积的液流速率随树干胸径的增大而减小。     

基于Holdridge和CCA分析的中国生态地理分区的比较

在前人工作基础上,对中国自然地理要素与生态地理区域的关系进行了综合分析,采用全国地形、土壤、气候、植被及遥感植被指数等数据,综合分析中国范围生态地理区域的分异规律,制订了生态地理分区的初步方案,并建立了相应的地理信息系统.基于Holdridge模型和CCA分析划分中国生态地理分区,建立了分区的指标体系,得到中国生态分区的大致界线,初步总结了各生态地理分区的地形、植被、气候等综合自然地理特征,完成对中国区域生态地理分区的划分.基于CCA分析的生态地理的分区,不仅结合自然区划和生态地理两种方法,而且加入了生态群落和遥感数据的综合应用.结果显示,由于受到模型适用性及数据误差的原因,基于CCA分析的结果比Holdridge模型的结果更合理一些.     

云南秋海棠属植物有性杂交特性

通过云南秋海棠属植物属下5个系统分类组的种类及外来园艺品种共109个组合的有性杂交试验,以系统分类组合、园艺分类组合(不同茎的形态类型组合)分别进行方差分析。结果表明:云南秋海棠属植物属下等级的系统分类组内、组间的有性杂交,以及不同茎的形态类型的有性杂交均无显著差异,亲和力较强而可育性高,可在云南秋海棠属植物属内进行广泛地远缘杂交; 云南产秋海棠属植物原种与外来园艺品种间的有性杂交亲和性弱而可育性低,需选择花粉粒和胚囊正常发育的外来园艺品种大量杂交,并采用切实可行的技术措施克服远缘杂交不育,以期培育新颖奇特的秋海棠属植物新品种。