Generalized Linear Dynamics of a Plant-Parasitic Nematode Population and the Economic Evaluation of Crop Rotations

In 1-year experiments, the final population density of nematodes is usually modeled as a function of initial density. Often, estimation of the parameters is precarious because nematode measurements, although laborious and expensive, are imprecise and the range in initial densities may be small. The estimation procedure can be improved by using orthogonal regression with a parameter for initial density on each experimental unit. In multi-year experiments parameters of a dynamic model can be estimated with optimization techniques like simulated annealing or Bayesian methods such as Markov chain Monte Carlo (MCMC). With these algorithms information from different experiments can be combined. In multi-year dynamic models, the stability of the steady states is an important issue. With chaotic dynamics, prediction of densities and associated economic loss will be possible only on a short timescale. In this study, a generic model was developed that describes population dynamics in crop rotations. Mathematical analysis showed stable steady states do exist for this dynamic model. Using the Metropolis algorithm, the model was fitted to data from a multi-year experiment on Pratylenchus penetrans dynamics with treatments that varied between years. For three crops, parameters for a yield loss assessment model were available and gross margin of the six possible rotations comprising these three crops and a fallow year were compared at the steady state of nematode density. Sensitivity of mean gross margin to changes in the parameter estimates was investigated. We discuss the general applicability of the dynamic rotation model and the opportunities arising from combination of the model with Bayesian calibration techniques for more efficient utilization and collection of data relevant for economic evaluation of crop rotations.     

Why ribonucleases induce tumor cell death

The characteristics and the possible mechanisms of action of cytotoxic ribonucleases (RNases), promising antitumor drugs, are described. Original experimental data and the results of analysis of recent publications have made it possible to identify the cellular components providing for the selective effects of exogenous RNases on tumor cells, on the one hand, and to estimate the contributions of individual molecular determinants to the enzyme cytotoxicity, on the other hand. The predominant effect of the electric charge of the RNase molecule on the induction of cell death has been demonstrated. The cytotoxic effects of RNases are determined by the catalytic cleavage of accessible RNA, the action of the products of its hydrolysis, and the noncatalytic electrostatic interaction of the exogenous enzyme with cell components. Potential RNase targets in a tumor cell and the role of modulation of calcium-dependent potassium channels and the ras oncogene in RNase-induced cell damage are considered. The effect of cytotoxic RNases on gene expression by affecting RNA interference is discussed.Translated from Molekulyarnaya Biologiya, Vol. 39, No. 1, 2005, pp. 3–13.Original Russian Text Copyright © 2005 by Ilinskaya, Makarov.     

嗜热拟青霉固体发酵产木聚糖酶的纯化和性质

研究一株新的嗜热拟青霉J18的固体发酵产木聚糖酶的纯化和性质。固体发酵的粗酶液经硫酸铵沉淀、凝胶过滤层析和离子交换层析得到了一种分子量约为26 kDa的电泳纯木聚糖酶,酶活力回收率为33.5%,纯化了5.27倍。该木聚糖酶具有很好的温度和pH稳定性,在pH7.0~pH 9.0下,60℃处理24 h,酶活力能保存80%以上。该酶水解玉米芯木聚糖生成以木二糖、木三糖和木四糖为主的低聚木糖,薄层层析分析表明不含木糖,适合生产低聚木糖。     

Annual cycle and inter-annual variability of gross primary production and ecosystem respiration in a floodprone river during a 15-year period

1. Temporal variation in ecosystem metabolism over a 15‐year period (1986–2000) was evaluated in a seventh order channelised gravel bed river (mean annual discharge 48.7 m³ s^?1) of the Swiss Plateau. The river is subject to frequent disturbance by bed‐moving spates. Daily integrals of gross primary production (GPP) and ecosystem respiration (ER) were calculated based on single‐station diel oxygen curves. 2. Seasonal decomposition of the time series of monthly metabolism rates showed that approximately 50% of the variation of GPP and ER can be attributed to season. Annual GPP averaged 5.0 ± 0.6 g O₂ m^?2 day^?1 and showed no long‐term trend. 3. Ecosystem respiration, averaging 6.2 ± 1.4 g O₂ m^?2 day^?1, declined from 8.8 to 4.1 g O₂ m^?2 day^?1 during the 15‐year period. This significant trend paralleled a decline in nitrate and soluble reactive phosphorus concentrations, and the biochemical oxygen demand discharged by sewage treatment facilities upstream of the study reach. The ratio of GPP to ER (P/R) increased from 0.53 to about 1 as consequence of ER reduction. 4. Bed moving spates reduced GPP by 49% and ER by 19%. Postspate recovery of GPP was rapid between spring and autumn and slow during winter. Recovery of ER lacked any seasonal pattern. Annual patterns of daily GPP and to a minor extent of daily ER can be described as a sequence of recovery periods frequently truncated by spates. 5. The study showed that disturbance by frequent bed‐moving spates resulted in major stochastic variation in GPP and ER but annual patterns were still characterised by a distinct seasonal cycle. It also became evident that stream metabolism is a suitable method to assess effects of gradual changes in water quality.     

Persistence and stability of macroinvertebrate communities in streams of Denali National Park, Alaska: implications for biological monitoring

1. Macroinvertebrate communities were studied from 1994 to 2001/2002 (except 1997) in six streams in Denali National Park, interior Alaska. All six streams were potential reference streams with no known impairment. 2. Abundance of individual taxa varied markedly from year to year. Overall, abundance decreased over the study period, particularly with respect to mayflies. Stonefly taxa showed lower persistence and were sometimes absent from a stream in any particular year. 3. Mean community persistence for the six streams, as measured by Jaccard's similarity coefficients between years, varied from 0.48 in the year pair 1999–2000 to 0.78 in 1998–99. Tattler Creek (a small stable stream) supported the most persistent macroinvertebrate community and Highway Pass Creek (a small, unstable creek) the least. Mean community persistence showed a significant relationship with mean winter snowfall (November to March) for the six streams. 4. The highest community compositional stability was found in Tattler Creek and the lowest in Highway Pass Creek, but stability varied markedly over time for the six streams, peaking in 1994–95 and reaching a minimum in 2000–01. Compositional stability was significantly related to the Pfankuch Index of channel stability. 5. The composition metrics % Chironomidae, % dominant taxa, %EPT, % Ephemeroptera and % Plecoptera, employed as part of the Alaska Stream Condition Index, varied over almost their entire range in these pristine streams across the 9 years of the study. 6. This study demonstrates the wide range of natural variation that occurs in benthic macroinvertebrate communities in these pristine central Alaskan streams, potentially limiting the applicability of composition metrics for the biological monitoring of water quality in these systems.     

Exogenous abscisic acid increases stability of polysomes in embryos of triticale caryopses during germination

Some posttranslational processes that occur in embryos of germinating triticale caryopses treated with different concentrations of abscisic acid (ABA) were examined. ABA increased the ratio of cytoskeleton-bound polysomes in the total population of polysomes and depressed the share of free and membrane-bound polysomes. Using exogenous RNase, stability of the total polysomal population as well as each polysomal fraction was investigated. The total extractable polysomes isolated from embryonic tissues of germinating triticale caryopses treated with ABA were more stable than the polysomes isolated from the control sample caryopses. The contribution of the polysomes that were not digested by RNase was increased by higher concentrations of ABA applied during germination. At high concentrations of ABA (50, 100 μM), the quantitative contribution of polysomes in the total ribosomal fraction was almost 100% of the amount of polysomes before digestion and the modifications observed consisted mainly of the shift of the so-called heavy polysomes towards light polysomes, containing a few ribosomes. Within each polysomal population, cytoskeleton-bound polysomes (CBP and CMBP) were the most stable, which may imply that the bonds between polysomes and these protein filaments, created in all eukaryotic cells increased their stability. It is assumed that mRNAs are stabilised or destabilised by interaction of proteins with their various sequences. A plant hormone may depress or elevate the quantities of these proteins, thus regulating the stability of different mRNAs. The results confirm the multi-faceted mechanism of ABA-induced response, where one of the constituents is the effect of ABA on the stability of mRNAs molecules. The co-ordinated regulation of mRNAs synthesis and their stability provide plants with improved adaptability.     

拮抗菌株的筛选及其发酵滤液抑菌活性考察

从土壤中筛选出3株具有抑菌活性的菌株,其中1#菌株对金黄色葡萄球菌、枯草芽孢杆菌、灰霉葡萄孢菌、变异链球菌、大肠杆菌均有较强抑制作用,初步鉴定为地衣芽孢杆菌。对其性质进行考察结果表明,在30℃,pH 7.0,150 r/min的条件下,恒温振荡培养24 h,测其发酵滤液的抑菌活力约为2 000 IU/mL。该发酵滤液具有较强的热稳定性,60℃放置1 h,抑菌活性下降了29%;100℃放置30 min,抑菌活性下降了45%,放置1 h,抑菌活性下降了48%;121℃放置1 h,抑菌活性仍能保持40%。该发酵滤液抑菌作用的最适pH值为7.0,在pH 3.0时能保持71%的抑菌活性,在pH 11.0时,仍能维持67%的抑菌活性,此发酵滤液对蛋白酶K不敏感,并且蛋白酶K对该发酵滤液的抑菌活性也起一定的促进作用。     

克服位置效应提高外源基因在CHO细胞中稳定高效表达的策略

能够生产有功用的治疗性蛋白的一个重要前提是获得稳定的重组蛋白高表达细胞株,然而筛选一个能够持续稳定表达外源蛋白的重组细胞株是费时费力的过程。有多篇文献报道了重组蛋白细胞株表达的不稳定性。位置效应是高表达细胞株不稳定性的重要因素,克服或利用位置效应是当前获得稳定高表达重组蛋白细胞株的有效途径。为解决外源基因插入的随机性所带来的不可预知的后果,可以事先在CHO细胞基因组中筛选转录热点区域,再通过位点特异性或同源重组的方式,实现外源基因的定点整合。各种调节位置效应的DNA元件陆续被发现,可以利用它们去调控基因表达及增加细胞株的稳定性。     

Study of the thermal stability of D-amino acid oxidase from Trigonopsis variabilis reveals enzyme inactivation via multiple steps

The thermal stability of a highly purified preparation of D-amino acid oxidase from Trigonopsis variabilis (TvDAO), which does not show microheterogeneity due to the partial oxidation of Cys-108, was studied based on dependence of temperature (20-60°C) and protein concentration (5-100 µmol L^-1). The time courses of loss of enzyme activity in 100 mmol L^-1 potassium phosphate buffer, pH 8.0, are well described by a formal kinetic mechanism in which two parallel denaturation processes, partial thermal unfolding and dissociation of the FAD cofactor, combine to yield the overall inactivation rate. Estimates from global fitting of the data revealed that the first-order rate constant of the unfolding reaction (k _a) increased 10⁴-fold in response to an increase in temperature from 20 to 60°C. The rate constants of FAD release (k _b) and binding (k _-b) as well as the irreversible aggregation of the apo-enzyme (k _agg) were less sensitive to changes in temperature, their activation energy (E _a) being about 52 kJ mol^-1 in comparison with an E _a value of 185 kJ mol^-1 for k _a. The rate-determining step of TvDAO inactivation switched from FAD dissociation to unfolding at high temperatures. The model adequately described the effect of protein concentration on inactivation kinetics. Its predictions regarding the extent of FAD release and aggregation during thermal denaturation were confirmed by experiments. TvDAO is shown to contain two highly reactive cysteines per protein subunit whose modification with 5,5'-dithio-bis (2-nitrobenzoic acid) was accompanied by inactivation. Dithiothreitol (1 mmol L^-1) enhanced up to 10-fold the recovery of enzyme activity during ion exchange chromatography of technical-grade TvDAO. However, it did not stabilize TvDAO at all temperatures and protein concentrations, suggesting that deactivation of cysteines was not responsible for thermal denaturation.     

Environmental Enhancement Through Use of Recycled Aggregate Concrete in a Two-Stage Mixing Approach

As a high priority of waste management and recycling by the Hong Kong government, Recycled Aggregate (RA) has been used in various construction applications, mainly as sub-grade, roadwork, and unbound materials. However, higher-grade applications are rare. The major barrier encountered is the variation of quality within RA, which causes lower strength, resulted from crystallization of Recycled Aggregate Concrete (RAC). Therefore, the objective of this study is to examine the crystallization of RAC in a Two-Stage Mixing Approach. Following are the five areas of interest: (i) investigate the waste problems in construction activities; (ii) examine the crystal development on the hydration of cement paste; (iii) develop a two-stage mixing approach (TSMA) for improving the performance of RAC; (iv) explore the crystallization of TSMA in comparison with the Normal Mixing Approach (NMA) through use of Differential Scanning Calorimetry (DSC); and (v) verify the results obtained from DSC analysis with those obtained from compressive strength testing. This study adopted 0, 20, and 100% RA substitution in virgin aggregate and measured by DSC and compressive strength on both TSMA and NMA. TSMA uses only half the water for mixing, forming a thin layer of cement slurry on the surface of RA that will permeate into the porous old cement mortar and fill old cracks and voids in the pre-mix process. The results from DSC analysis clearly demonstrated that TSMA can give a better crystallization of CaO·SiO₂·H₂O [CSH] and Ca(OH)₂[CH]. The optimal situation occurs on 20% RA substitution in virgin aggregate, balancing the advantages of each, a finding supported by the results from compressive strength testing. Therefore, TSMA is a superior methodology and opens a wider application for the use of RAC.