Testing ant predation on the coffee berry borer in shaded and sun coffee plantations in Colombia

Soil‐dwelling ants, many of which are generalist predators, are more diverse in shaded than in sun coffee plantations without trees. We compared ant predation on the coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae: Scolytinae) in three shaded and three sun coffee plantations in Apía, Colombia, in both the wet and the dry seasons. We found that H. hampei adults exposed to ants for 5 days suffered higher removal in shaded plantations and in the wet season. In the laboratory, we observed that ants killed 74–99% of H. hampei adults over the course of 5 days. Ants appear to be important predators of H. hampei, particularly in shaded coffee plantations and in the wet season.     

An approach for post‐market monitoring of potential environmental effects of Bt‐maize expressing Cry1Ab on natural enemies

Post‐market monitoring (PMM) consistent with Swiss and European Union legislation should ensure the detection and prevention of adverse effects on the environment possibly deriving from commercial cultivation of genetically modified (GM) crops. Insect‐resistant GM crops (such as Bt‐maize) raise particular questions regarding disturbances of biological control functions provided by beneficial insects such as predators and parasitoids (so‐called natural enemies). Consensus among regulators, scientists and the agricultural biotech industry on appropriate PMM plans allowing the detection and possibly prevention of such adverse effects is still lacking. The aims of this study were to identify the necessity for PMM of Bt‐maize expressing Cry1Ab on natural enemies and to develop an appropriate PMM plan. The approach chosen consisted in determining what type of monitoring is most appropriate to address potential effects of Bt‐maize on natural enemies during commercial cultivation. This included identifying whether there remain substantial scientific uncertainties that would support case‐specific monitoring. Existing pre‐market risk assessment data indicate that Bt‐maize (Cry1Ab) comprises a negligible risk for disturbances in biological control functions of natural enemies. As a consequence, a faunistic monitoring of specific groups of natural enemies is not considered an appropriate approach to detect failures in biological control functions. Alternatively, an approach is proposed that consists in indirectly analysing biological control functions by surveying outbreaks of maize herbivores. Unusual herbivore outbreaks could indicate failures in biological control functions of natural enemies. Data could be collected via questionnaires addressed to farmers growing Bt‐maize. Significant correlations between unusual occurrences of specific maize herbivores and the cultivation of Bt‐maize would subsequently need specific studies to determine possible causalities in more detail. The here proposed approach has the advantage of covering different natural enemy groups. It represents a cost‐effective strategy to obtain scientifically sound data as a basis for regulatory decision‐making.     

Molecular genetic variation and geographical structuring in Fusarium graminearum

Inter‐simple sequence repeat markers were employed to study the genetic structure of Fusarium graminearum populations collected from three Canadian provinces. Our study suggested high genetic diversity and frequent gene flow among population samples analysed. The analysis of molecular variance indicated that most of the gene diversity (91.78% to 97.23%) was distributed within populations. Frequent gene flow among the western prairie provinces and cluster analysis results indicated that the population sample from Alberta was closer to Saskatchewan than Manitoba, which could be the result of movement of the pathogen via infected grain or through wind‐borne ascospore dispersal. Analysis of multilocus associations showed that all populations were in linkage equilibrium, indicating that sexual recombination is a frequent phenomenon in F. graminearum populations from western Canada.     

Minimal sample requirement for highly multiplexed protein quantification in cell lines and tissues by PCT‐SWATH mass spectrometry

The amount of sample available for clinical and biological proteomic research is often limited and thus significantly restricts clinical and translational research. Recently, we have integrated pressure cycling technology (PCT) assisted sample preparation and SWATH‐MS to perform reproducible proteomic quantification of biopsy‐level tissue samples. Here, we further evaluated the minimal sample requirement of the PCT‐SWATH method using various types of samples, including cultured cells (HeLa, K562, and U251, 500 000 to 50 000 cells) and tissue samples (mouse liver, heart, brain, and human kidney, 3–0.2 mg). The data show that as few as 50 000 human cells and 0.2–0.5 mg of wet mouse and human tissues produced peptide samples sufficient for multiple SWATH‐MS analyses at optimal sample load applied to the system. Generally, the reproducibility of the method increased with decreasing tissue sample amounts. The SWATH maps acquired from peptides derived from samples of varying sizes were essentially identical based on the number, type, and quantity of identified peptides. In conclusion, we determined the minimal sample required for optimal PCT‐SWATH analyses, and found smaller sample size achieved higher quantitative accuracy.     

On‐line yeast propagation process monitoring and control using an intelligent automatic control system

The monitoring and control of bioprocesses is a challenging task. This applies particularly if the actions to the process have to be carried out in real‐time. This work presents a system for on‐line monitoring and control of batch yeast propagation under limiting conditions based on a virtual plant operator, which uses the concept of intelligent control algorithms by means of fuzzy logic theory. Process information is provided on‐line using a sensor array comprising the measurement of OD, operating temperature, pressure, density, dissolved oxygen, and pH value. In this context practical problems arising through on‐line sensing and signal processing are addressed. The preprocessed sensor data are fed to a neural network for on‐line biomass estimation. The root mean squared error of prediction is 4 × 10⁶ cells/mL. The proposed system then triggers temperature and aeration by usage of a temperature dependent metabolic growth model and sensor data. The deviation of the predicted biomass from that of the reference trajectory as modeled by the metabolic growth model and its temporal derivative are used as inputs for the fuzzy temperature controller. The inputs used by the fuzzy aeration controller are the deviation of measured extract from that of the reference trajectory, the predicted cell count, and the dissolved oxygen concentration. The fuzzy‐based expert system allows to provide the desired yeast cell concentration of 100–120 × 10⁶ cells/mL at a minimum residual extract limit of 6.0 g/100 g at the required point of time. Thus, a dynamic adjustment of the propagation process to the overall production schedule is possible in order to produce the required amount of biomass at the right time.     

LAMP-LFD技术及其在生物快检方面应用

LAMP-LFD技术是环介导横温扩增技术(Loop-mediated isothermal amplification)与横向流动试纸条(Lateral flow dipstick)相结合的一种新颖的检测技术,该技术将核酸横温扩增和可视化试纸条检测有机地结合。环介导等温扩增技术(LAMP),只需要在恒温(60℃~65℃)条件下保温几十分钟,即可将所需目的基因扩增到109水平。横向流动试纸条(LFD),融合了免疫层析技术和分子生物学手段,能在纸条上形成有颜色的检测线从而可以特异性地检测出该目标产物。环介导横温扩增技术(LAMP)与横向流动试纸条(LFD)相互结合的技术,使LAMP扩增产物现场检测可视化,检测结果明显直观,通过肉眼即可辨认,并具有高特异性、高灵敏度、简便、安全及成本低的特点,一度引起科研工作者的广泛关注。综述了环介导横温扩增方法与横向流动试纸条相结合(LAMP-LFD)技术的原理、优势、及其在生物快速检测方面的应用等,并指出了LAMP-LFD技术目前存在的不足以及展望。     

抑癌基因PTEN转基因小鼠的构建及表型初步分析

目的:构建抑癌基因PTEN(phosphatase and tensin homolog deleted on chromosome ten)的转基因小鼠模型并对其表型进行初步分析。方法:细菌人工染色体(BAC)载体系统构建打靶载体创建PTEN转基因小鼠模型;利用对鼠尾DNA进行PCR检测的方法对出生的F0代小鼠进行基因型鉴定,将阳性F_0代小鼠与野生型小鼠交配繁殖筛选稳定遗传的转基因系。分离并培养小鼠胚胎成纤维细胞(mouse embryo fibroblast,MEF),利用Western blotting检测比较转基因阳性小鼠与同窝野生型小鼠MEF细胞中PTEN的蛋白表达水平,并通过克隆形成实验对比PTEN转基因小鼠与野生型小鼠MEF细胞的增殖能力;取成年小鼠主要组织器官提取蛋白,Western blotting检测PTEN转基因小鼠主要组织的PTEN蛋白表达情况;从小鼠出生后第三周开始统计、分析并制作小鼠的体重生长曲线;此外,还对比了PTEN转基因小鼠与野生型小鼠肺、肝、脾脏的细胞大小与腹腔内的脂肪含量。结论:PTEN转基因小鼠能够存活并稳定遗传;Western blotting结果表明,不论在胚胎期还是成年期,PTEN转基因小鼠体内的PTEN蛋白水平均高于同窝的野生型小鼠,转基因小鼠的PTEN表达水平接近野生型水平的3倍;对PTEN转基因小鼠的整体表型进行初步分析,发现Pten基因在体内过表达后,小鼠的体型显著变小,而细胞大小不变;腹腔内的脂肪含量显著减少。结论:成功构建了PTEN转基因小鼠模型,并获得了生理条件下PTEN过表达的原代细胞系,为研究抑癌基因PTEN的体内生理功能提供了重要的动物模型。     

Efficacy of Metarhizium anisopliae in controlling the two‐spotted spider mite Tetranychus urticae on common bean in screenhouse and field experiments

The efficacy of aqueous and emulsifiable formulations of the fungus Metarhizium anisopliae isolate ICIPE78 was evaluated on the population density of Tetranychus urticae infesting common bean plants under screenhouse and field conditions. Synthetic acaricide abamectin was included as a check. Bean plants were artificially infested with T. urticae and allowed to multiply. Three treatments were applied in the screenhouse and 1 treatment in field trials. Mite density was recorded 2 d before spraying and weekly postspraying. The number of pods per plant, number of seeds per pod, and the dry weight of seeds per plant were recorded only in the screenhouse trials. In both screenhouse and field trials, fungal formulations applied at the concentration of 10⁸ conidia/mL and the acaricide reduced the population density of mites as compared to the controls. There were significant differences in T. urticae population densities between the treatments at the various post‐spraying sampling dates. In the screenhouse, the mite densities were near zero from 3‐week postspraying in the treated leaves. At 4‐week postspraying, there were no more leaves in the untreated control (T1) and in the control water + Silwet‐L77 (T2). Fungal formulations were as effective as abamectin in reducing mite densities in both screenhouse and field experiments. There were significant differences in the production parameters during the 2 screenhouse trials, with fungal and abamectin treatments generally having the highest yield. Results of this study underline the potential of the M. anisopliae isolate ICIPE78 as an alternative to acaricides for T. urticae management.     

两种高危型HPV检测技术诊断宫颈病变的临床价值比较

目的:比较酶切信号放大法(Cervista)与导流杂交基因芯片技术(Hybri Ma)检测高危型人乳头状瘤病毒(HR-HPV)诊断宫颈上皮内瘤变2级或2级以上(CIN2+)的临床价值。方法:随机选择288例2012年3月至2013年1月在哈尔滨医科大学附属第一医院妇科门诊进行新柏氏液基细胞学检查的年龄在20~65岁的宫颈细胞学检测未明确意义的不典型鳞状细胞(ASCUS)的患者,采用Cervista技术与Hybri Ma技术进行高危型HPV检测,并对入组的患者行阴道镜下宫颈活组织检查。以病理学诊断结果为金标准,比较Cervista技术与Hybri Ma技术诊断宫颈上皮内瘤变2级或2级以上(CIN2+)的敏感度、特异度及ROC曲线。结果:在入组的288例患者中,Cervista技术和Hybri Ma技术检出高危型HPV的阳性率分别为49.31%和51.39%(P0.05),其诊断CIN2+的敏感度分别为95.65%和91.30%(P0.05),特异度分别为59.50%和56.20%(P0.05),阳性预计值分别为30.99%和28.38%(P0.05),阴性预计值分别为98.63%和97.14%(P0.05)。两组ROC曲线下面积分别为0.776和0.738(P0.05)。结论:Cervista技术与Hybri Ma技术诊断CIN2+的临床价值相当。