Predicting variability in biological control of a plant-pathogen system using stochastic models.

A stochastic model for the dynamics of a plant-pathogen interaction is developed and fitted to observations of the fungal pathogen Rhizoctonia solani (Kühn) in radish (Raphanus sativus L.), in both the presence and absence of the antagonistic fungus Trichoderma viride (Pers ex Gray). The model incorporates parameters for primary and secondary infection mechanisms and for characterizing the time-varying susceptibility of the host population. A parameter likelihood is developed and used to fit the model to data from microcosm experiments. It is shown that the stochastic model accounts well for observed variability both within and between treatments. Moreover, it enables us to describe the time evolution of the probability distribution for the variability among replicate epidemics in terms of the underlying epidemiological parameters for primary and secondary infection and decay in susceptibility. Consideration of profile likelihoods for each parameter provides strong evidence that T. viride mainly affects primary infection. By using the stochastic model to study the dependence of the probability distribution of disease levels on the primary infection rate we are therefore able to predict the effectiveness of a widely used biological control agent.     

Transmission rates and adaptive evolution of pathogens in sympatric heterogeneous plant populations

Diversification in agricultural cropping patterns is widely practised to delay the build-up of virulent races that can overcome host resistance in pathogen populations. This can lead to balanced polymorphism, but the long-term consequences of this strategy for the evolution of crop pathogen populations are still unclear. The widespread occurrence of sibling species and reproductively isolated sub-species among fungal and oomycete plant pathogens suggests that evolutionary divergence is common. This paper develops a mathematical model of host-pathogen interactions using a simple framework of two hosts to analyse the influences of sympatric host heterogeneity on the long-term evolutionary behaviour of plant pathogens. Using adaptive dynamics, which assumes that sequential mutations induce small changes in pathogen fitness, we show that evolutionary outcomes strongly depend on the shape of the trade-off curve between pathogen transmission on sympatric hosts. In particular, we determine the conditions under which the evolutionary branching of a monomorphic into a dimorphic population occurs, as well as the conditions that lead to the evolution of specialist (single host range) or generalist (multiple host range) pathogen populations.     

Comparative losses of quantitative and molecular genetic variation in finite populations of Drosophila melanogaster

Quantitative genetic variation, the main determinant of the ability to evolve, is expected to be lost in small populations, but there are limited data on the effect, and controversy as to whether it is similar to that for near neutral molecular variation. Genetic variation for abdominal and sternopleural bristle numbers and allozyme heterozygosity were estimated in 23 populations of Drosophila melanogaster maintained at effective population sizes of 25, 50, 100, 250 or 500 for 50 generations, as well as in 19 highly inbred populations and the wild outbred base population. Highly significant negative regressions of proportion of initial genetic variation retained on inbreeding due to finite population size were observed for both quantitative characters (b = -0.67 +/- 0.14 and -0.58 +/- 0.11) and allozyme heterozygosity (b = -0.79 +/- 0.10), and the regression coefficients did not differ significantly. Thus, quantitative genetic variation is being lost at a similar rate to molecular genetic variation. However, genetic variation for all traits was lost at rates significantly slower than predicted by neutral theory, most likely due to associative overdominance. Positive, but relatively low correlations were found among the different measures of genetic variation, but their low magnitudes were attributed to large sampling errors, rather than differences in the underlying processes of loss.     

A bacterial cell to cell signal in the lungs of cystic fibrosis patients

Pseudomonas aeruginosa is an opportunistic pathogen that is a major cause of mortality in cystic fibrosis (CF) patients. This bacterium has numerous genes controlled by cell to cell signaling, which occurs through a complex circuitry of interconnected regulatory systems. One of the signals is the Pseudomonas Quinolone Signal (PQS), which was identified as 2-heptyl-3-hydroxy-4-quinolone. This intercellular signal controls the expression of multiple virulence factors and is required for virulence in an insect model of P. aeruginosa infection. Previous studies have implied that the intercellular signals of P. aeruginosa are important for human disease, and our goal was to determine whether PQS was produced during human infections. In this report, three types of samples from CF patients infected with P. aeruginosa were analyzed for the presence of PQS. Sputum, bronchoalveolar lavage fluid, and mucopurulent fluid from distal airways of end-stage lungs removed at transplant, all contained PQS, indicating that this cell to cell signal is produced in vivo by P. aeruginosa infecting the lungs of CF patients.     

Cost-Effective Control of Plant Disease When Epidemiological Knowledge Is Incomplete: Modelling Bahia Bark Scaling of Citrus

A spatially-explicit, stochastic model is developed for Bahia bark scaling, a threat to citrus production in north-eastern Brazil, and is used to assess epidemiological principles underlying the cost-effectiveness of disease control strategies. The model is fitted via Markov chain Monte Carlo with data augmentation to snapshots of disease spread derived from a previously-reported multi-year experiment. Goodness-of-fit tests strongly supported the fit of the model, even though the detailed etiology of the disease is unknown and was not explicitly included in the model. Key epidemiological parameters including the infection rate, incubation period and scale of dispersal are estimated from the spread data. This allows us to scale-up the experimental results to predict the effect of the level of initial inoculum on disease progression in a typically-sized citrus grove. The efficacies of two cultural control measures are assessed: altering the spacing of host plants, and roguing symptomatic trees. Reducing planting density can slow disease spread significantly if the distance between hosts is sufficiently large. However, low density groves have fewer plants per hectare. The optimum density of productive plants is therefore recovered at an intermediate host spacing. Roguing, even when detection of symptomatic plants is imperfect, can lead to very effective control. However, scouting for disease symptoms incurs a cost. We use the model to balance the cost of scouting against the number of plants lost to disease, and show how to determine a roguing schedule that optimises profit. The trade-offs underlying the two optima we identify—the optimal host spacing and the optimal roguing schedule—are applicable to many pathosystems. Our work demonstrates how a carefully parameterised mathematical model can be used to find these optima. It also illustrates how mathematical models can be used in even this most challenging of situations in which the underlying epidemiology is ill-understood.     

尿毒症皮肤瘙痒患者采取腹膜透析与血液透析治疗的效果探究

目的：探究尿毒症皮肤瘙瘁患者采取腹膜透析与血液透析治疗的临床效果,并为该病的临床治疗提供经验积累。方法：选取我院肾内科于2005年1月-2012年12月收治的52例尿毒症皮肤瘙痒患者,利用随机数字表法进行分组,分别设为研究组和对照组。其中对照组实施血液透析治疗,而研究组开展腹膜透析治疗。记录两组在治疗前和治疗后第8周末血生化客观指标及皮肤瘙痒程度,并做好对比。结果：两组在治疗前的皮肤瘙痒评分、β2-MG、PTH、p3,BUN、SCr值差异无统计学意义（P〉0．05）;治疗后,研究组皮肤瘙瘁评分为（3．4±0．8）分,对照组为（5．8±0．9）分,差异有统计学意义（P〈0．05）。治疗后,研究组β2-MG、PTH及Ps．均低于对照组,差异有统计学意义（P〈0．05）。结论：尿毒症皮肤瘙痒患者的主要致敏因子为大中分子,采取腹膜透析能够有效清除大分子物质,进而达到瘙痒程度的有效改善。     

丝素蛋白/羟基磷灰石复合物对磷酸钙骨水泥性能的影响

目的：磷酸钙骨水泥（Calcium phosphate cement,CPC）以其诸多优点正得到了越来越多的应用,但其较差的力学性能表现也限制了它的使用范围。本研究目的在于改善磷酸钙骨水泥的力学性能,同时评估改性后的磷酸钙骨水泥的其他性能。方法：通过丝素蛋白（Silk fibroin,SF）的矿化自组装方法制备丝素蛋白／羟基磷灰石复合物（silk fibroin/hydroxyapitite composite, SF/HA）。按照1％、2％、3％、4％的质量分数加入磷酸钙骨水泥中,与磷酸钙骨水泥组对比。比较内容包括力学强度、抗渍散性能及细胞毒性。结果：以丝素蛋白溶液为液相组的磷酸钙骨水泥强度大约为35MPa。随后随着添加丝素蛋白／羟基磷灰石复合物的质量分数从1％增至3％,磷酸钙骨水泥的强度逐渐增加（P〈0．05）,最高约至45MPa。而当丝素蛋白／羟基磷灰石的质量分数达到4％时,磷酸钙骨水泥的强度较质量分数3％组小幅度下降至43MPa（P〈0．05）。以丝素蛋白溶液作为液相时,磷酸钙骨水泥的抗溃散能力也得到了加强。在MTT法测定细胞活力的对照实验中,无论是加入丝素蛋白溶液或丝素蛋白／羟基磷灰石复合物,都未观察到细胞毒性。结论：在磷酸钙骨水泥中加入3％质量分数的丝素蛋白／羟基磷灰石复合物,能显著提高磷酸钙骨水泥的抗压强度。而丝素蛋白溶液作为液相可改善磷酸钙骨水泥的抗溃散能力。同时,丝素蛋白和丝素蛋白／羟基磷灰石复合物都不表现出细胞毒性。更理想的力学强度和更强的抗溃散能力,大大扩展了磷酸钙骨水泥的应用范围。     

重组人MASP2蛋白在大肠杆菌中的表达和纯化

目的:获得酶原形式的重组人甘露聚糖结合凝集素相关丝氨酸蛋白酶2(MASP2)。方法:在大肠杆菌中诱导表达重组人MASP2全长蛋白,包涵体裂解后,经复性、透析、浓缩、考马斯亮蓝染色、SDS-PAGE及Western印迹,鉴定纯化结果及酶活性。结果:复性后的MASP2蛋白经考马斯亮蓝染色未见杂带。自激活实验表明,当MASP2浓度在1μmool/L以下时,无论在4℃还是37℃,都能较稳定地保持酶原形式;蛋白浓度为3.5μmool/L时只能在4℃保持稳定,37℃发生自激活;蛋白浓度达到12μmool/L后,在4℃时已不能稳定存在。结论:获得了较纯的重组人MASP2蛋白,且具有自激活活性。     

变性高效液相色谱技术对创伤弧菌检测的研究

应用PCR结合变性高效液相色谱技术对创伤弧菌进行检测,建立创伤弧菌快速准确的检测新方法。经过DHPLC分析条件优化,在DHPLC非变性温度下分析创伤弧菌特异性PCR扩增产物。同时进行方法特异性、灵敏度、重复性实验。实验结果表明所建立的创伤弧菌PCR-DHPLC检测方法特异性强、灵敏度高、重现性好、结果稳定可靠、检测时间短,检测低限可达到124 CFU/mL,是创伤弧菌快速检测的新技术。