Mini-scale bioprocessing systems for highly parallel animal cell cultures

Animal cells have been used extensively in therapeutic protein production. The growth of animal cells and the expression of therapeutic proteins are highly dependent on the culturing environments. A large number of experimental permutations need to be explored to identify the optimal culturing conditions. Miniaturized bioreactors are well suited for such tasks as they offer high-throughput parallel operation and reduce cost of reagents. They can also be automated and be coupled to downstream analytical units for online measurements of culture products. This review summarizes the current status of miniaturized bioreactors for animal cell cultivation based on the design categories: microtiter plates, flasks, stirred tank reactors, novel designs with active mixing, and microfluidic cell culture devices. We compare cell density and product titer, for batch or fed-batch modes for each system. Monitoring/controlling devices for engineering parameters such as pH, dissolved oxygen, and dissolved carbon dioxide, which could be applied to such systems, are summarized. Finally, mini-scale tools for process performance evaluation for animal cell cultures are discussed: total cell density, cell viability, product titer and quality, substrates, and metabolites profiles.     

废水自养生物脱氮技术研究进展

基于短程硝化和厌氧氨氧化的自养脱氮工艺是生物脱氮领域研究的热点,它的发现为低碳氮比废水的处理提供了新的思路。近些年来,人们陆续开发了SHARON、ANAMMOX、CANON、OLAND等自养生物脱氮工艺,进一步推动了高效、低耗脱氮技术的开发和研究。本文从工艺原理、特点等方面,对自养生物脱氮工艺的国内外研究状况进行了总结和对比,并提出了存在的问题及发展方向。     

海洋碳迁移转化与主要化学驱动因子的相互关系

分析了化学驱动因子对海洋碳迁移转化过程的影响.海洋碳迁移转化与各种化学驱动因子参与的生物地球化学过程密切相关.营养盐水平、pH、溶解氧浓度（DO）、氧化还原电位（Eh）、SO₄^2-及硫电位（Es）等主要化学驱动因子的消长导致了海洋化学环境的变化,进而对海洋碳的迁移转化产生影响.在营养盐的供给和生物吸收情况良好的海域,CO₂由于光合作用,并通过沉降有机物的氧化,不断被转移到海水深层,使得海水中的CO₂分压（P_CO2）降低,CO₂的海-气交换量和有机碳输出通量增大,从使该海域表现为CO₂的汇.由于CO₂的溶解与吸收以及有机物的降解造成了海洋环境的日益酸化,引起了海水中碳酸盐溶解度增大;沉积物中酸碱环境的变化也与有机物的矿化以及碳酸盐的溶解、沉淀过程密切相关.此外,DO、Eh、SO₄^2-及Es的变化与水体中有机碳的矿化分解过程和碳在沉积层中沉积埋葬过程相耦合.在水体中,高DO、高Eh利于有机碳向无机碳转化;而在DO和Eh较低的沉积环境中,高SO₄^2-不利于有机碳的埋葬与保存.     

随机区块法在空间点格局分析中的应用

利用一种以随机模拟技术为基础的空间点格局分析方法--随机区块法对长白山阔叶红松林中5个主要树种的空间格局及空间关系进行了分析.随机区块法利用ICS(t)=s2(t)/x(t)-1来衡量不同尺度t上空间点的分布格局,并通过随机模拟技术构建零假设的95%或99%置信区间,以确定空间点格局偏离零假设的显著性程度.结果表明,除蒙古栎、山荆子、怀槐为随机分布外,其它树种均在局部空间尺度上呈聚集分布;并且绝大多数种对或种组在局部空间尺度上呈正相关关系.随机区块法克服了小样方统计法研究尺度单一的问题,也避免了以空间点距离为基础的点格局分析过程中边缘效应带来的影响和误差,尤其是在多变量点格局分析中具有明显的优势.     

Early-stage litter decomposition and its influencing factors in the wetland of the Sanjiang Plain, China

Using the litter bag technique, the decomposition rates and their influencing factors were studied by investigating three wetland macrophytes, Calamagrostics angustifolia, Carex meyeriana and Carex lasiocapa, in Sanjiang Plain, Northeast China. It was revealed that C. lasiocapa lost 28.91% of its weight, C. angustifolia lost 31.98% and C. meyeriana lost 32.99% after 164 days. Another finding was that the amount of organic carbon in the litter of C. angustifolia and C. lasiocapa fluctuated, but continuously decreased in that of C. meyeriana. However, all the three types of litter released organic carbon. Nitrogen was released substantially from the litter of both C. angustifolia and C. meyeriana, but accumulated in the litter of C. lasiocapa. Phosphorus concentrations in all the three types of litter apparently decreased first and then slightly increased. Overall, P release was observed in all the three types of litter. The C/N and C/P ratios varied significantly in the decomposition process. The decomposition rates and nutrient content variations were simultaneously influenced by the quality of the litter as well as the environmental factors in the Sanjian Plain, but they were more strongly affected by the quality of the litter.     

玉米秸秆分批补料获得高还原糖浓度酶解液的条件优化

木质纤维素高浓度还原糖水解液的获得是纤维乙醇产业化发展的方向。在发酵工业领域,分批补料法是实现这一目标的重要研究途径。本研究采用分批补料法对获得高浓度玉米秸秆酶解还原糖的条件进行了优化。以稀硫酸预处理的玉米秸秆为原料,考察了液固比、补加量与补加时间对分批补料糖化的影响。结果表明,秸秆高浓度酶解液条件的初始物料为20％ (重量/体积),木聚糖酶220 U/g (底物),纤维素酶6 FPU/g (底物),果胶酶50 U/g (底物),在24 h、48 h后分批补加8％预处理后的物料,同时添加与补料量相应的木聚糖酶20 U/g (底物),纤维素酶2 FPU/g (底物),72 h后,最终糖化结果与非补料法相比,还原糖浓度从48.5 g/L提高到138.5 g/L,原料的酶解率最终达到理论值的62.5％。试验结果表明补料法可以显著提高秸秆水解液还原糖浓度。     

A general comparison of relaxed molecular clock models

Several models have been proposed to relax the molecular clock in order to estimate divergence times. However, it is unclear which model has the best fit to real data and should therefore be used to perform molecular dating. In particular, we do not know whether rate autocorrelation should be considered or which prior on divergence times should be used. In this work, we propose a general bench mark of alternative relaxed clock models. We have reimplemented most of the already existing models, including the popular lognormal model, as well as various prior choices for divergence times (birth-death, Dirichlet, uniform), in a common Bayesian statistical framework. We also propose a new autocorrelated model, called the "CIR" process, with well-defined stationary properties. We assess the relative fitness of these models and priors, when applied to 3 different protein data sets from eukaryotes, vertebrates, and mammals, by computing Bayes factors using a numerical method called thermodynamic integration. We find that the 2 autocorrelated models, CIR and lognormal, have a similar fit and clearly outperform uncorrelated models on all 3 data sets. In contrast, the optimal choice for the divergence time prior is more dependent on the data investigated. Altogether, our results provide useful guidelines for model choice in the field of molecular dating while opening the way to more extensive model comparisons.     

Biofilm stratification during simultaneous nitrification and denitrification (SND) at a biocathode

The aeration of the cathode compartment of bioelectrochemical systems (BESs) was recently shown to promote simultaneous nitrification and denitrification (SND). This study investigates the cathodic metabolism under different operating conditions as well as the structural organization of the cathodic biofilm during SND. Results show that a maximal nitrogen removal efficiency of 86.9 ± 0.5%, and a removal rate of 3.39 ± 0.08 mg N L⁻¹ h⁻¹ could be achieved at a dissolved oxygen (DO) level of 5.73 ± 0.03 mg L⁻¹ in the catholyte. The DO levels used in this study are higher than the thresholds previously reported as detrimental for denitrification. Analysis of the cathodic half-cell potential during batch tests suggested the existence of an oxygen gradient within the biofilm while performing SND. FISH analysis corroborated this finding revealing that the structure of the biofilm included an outer layer occupied by putative nitrifying organisms, and an inner layer where putative denitrifying organisms were most dominant. To our best knowledge this is the first time that nitrifying and denitrifying microorganisms are simultaneously observed in a cathodic biofilm.     

Overcoming the thermodynamic limitation in asymmetric hydrogen transfer reactions catalyzed by whole cells

Whole lyophilized cells of an Escherichia coli overexpressing the alcohol dehydrogenase (ADH-'A') from Rhodococcus ruber DSM 44541 were used for the asymmetric reduction of ketones to secondary alcohols. The recycling of the required nicotinamide cofactor (NADH) was achieved in a coupled-substrate process. In the course of the reaction the ketone is reduced to the alcohol and the hydrogen donor 2-propanol is oxidized to acetone by one enzyme. This leads to a thermodynamic equilibrium between all four components determining the maximum achievable conversion. To overcome this limitation an in situ product removal technique (ISPR) for the application with whole cells was developed. In this method the most volatile compound is separated from the reaction vessel by an air flow resulting in a shift of the equilibrium towards the desired secondary alcohol. The so-called stripping process represents a simple and efficient method to overcome the thermodynamic limitation in biocatalytic reactions. Employing this method, the conversion of selected biotransformations was increased up to completeness.     

An approach to the residence time distribution for stochastic multi-compartment models

Stochastic compartmental models are widely used in modeling processes such as drug kinetics in biological systems. This paper considers the distribution of the residence times for stochastic multi-compartment models, especially systems with non-exponential lifetime distributions. The paper first derives the moment generating function of the bivariate residence time distribution for the two-compartment model with general lifetimes and approximates the density of the residence time using the saddlepoint approximation. Then, it extends the distributional approach to the residence time for multi-compartment semi-Markov models combining the cofactor rule for a single destination and the analytic approach to the two-compartment model. This approach provides a complete specification of the residence time distribution based on the moment generating function and thus facilitates an easier calculation of high-order moments than the approach using the coefficient matrix. Applications to drug kinetics demonstrate the simplicity and usefulness of this approach.