RNA assay as a method of viable biomass determination in the organic fraction of municipal solid waste suspension

A method of RNA determination by means of alkaline hydrolysis and UV detection was applied to evaluate the amount of viable biomass during the aerobic biodegradation of the organic fraction of the municipal solid waste suspension. The relative standard deviation of this determination was from 0.7 to 4%. The viable biomass constituted only 9% of the volatile suspended solids in the activated sludge.     

樟叶越桔细胞悬浮培养条件的优化

为了提高樟叶越桔(Vaccinium dunalianum)悬浮培养细胞的生物量, 以樟叶越桔叶片愈伤组织为试材, 通过单因素试验探究不同蔗糖浓度、培养基pH值、培养基体积、初始接种量和摇床转速对悬浮培养细胞生长的影响, 并根据响应面法Box-Behnken试验设计原理进行组合试验以优化培养条件。结果显示, 以改良WPM培养基为基础培养基, 樟叶越桔细胞悬浮培养的最优条件为40 g·L^-1蔗糖、培养基pH5.2、培养基体积45 mL、初始接种量2.64 g和摇床转速为149 r·min^-1, 其细胞生物量干重为0.184 4 g, 与理论预测值0.184 5 g较为接近, 且细胞的生长曲线呈S型。研究结果为樟叶越桔悬浮培养细胞次生代谢产物的生产调控奠定了技术基础。     

Effective conductivity of a suspension of permeabilized cells: a theoretical analysis

During the electroporation cell membrane undergoes structural changes, which increase the membrane conductivity and consequently lead to a change in effective conductivity of a cell suspension. To correlate microscopic membrane changes to macroscopic changes in conductivity of a suspension, we analyzed the effective conductivity theoretically, using two different approaches: numerically, using the finite elements method; and analytically, by using the equivalence principle. We derived the equation, which connects membrane conductivity with effective conductivity of the cell suspension. The changes in effective conductivity were analyzed for different parameters: cell volume fraction, membrane and medium conductivity, critical transmembrane potential, and cell orientation. In our analysis we used a tensor form of the effective conductivity, thus taking into account the anisotropic nature of the cell electropermeabilization and rotation of the cells. To determine the effect of cell rotation, as questioned by some authors, the difference between conductivity of a cell suspension with normally distributed orientations and parallel orientation was also calculated, and determined to be <10%. The presented theory provides a theoretical basis for the analysis of measurements of the effective conductivity during electroporation.     

Conductimetric assessment of the biomass content in suspensions of immobilised (gel-entrapped) microorganisms

Summary The problem of obtaining a rapid estimate of the microbial content of an immobilised cell suspension is addressed. The low-frequency conductivity of free-living cell suspensions of Clostridium pasteurianum is lower than that of the medium in which they are suspended, by an amount conforming to the Bruggeman relation. The conductivity of the cell wall makes a negligible contribution to the measured conductivity under the conditions used. Calcium alginate beads (lacking microbial cells) lower the conductivity of a solution with which they have been equilibrated by an extent which is a function of the concentration of alginate gel used in forming the beads. When this is taken into account, the ratio of the conductivity of a suspension of gel-immobilised cells to that of the suspending medium can be used to give a rapid and convenient assessment of the amount of microbial biomass present.     

Non-invasive methods for determination of cellular growth inPodophyllum hexandrum suspension cultures

Culture conductivity and on-line NADH fluorescence were used to measure cellular growth in plant cell suspension cultures ofPodophyllum hexandrum. An inverse correlation between dry cell weight and medium conductivity was observed during shake flask cultivation. A linear relationship between dry cell weight and culture NADH fluorescence was obtained during the exponential phase of batch cultivation in a bioreactor under the pH stat (pH 6) conditions. It was observed that conductivity measurement were suitable for biomass characterisation under highly dynamic uncontrolled shake flask cultivation conditions. However, if the acid/alkali feeding is done for pH control the conductivity measurement could not be applied. On the other hand the NADH fluorescence measurement allowed online-in situ biomass monitoring of rather heterogenous plant cell suspension cultures in bioreactor even under the most desirable pH stat conditions.     

Dielectric properties of synaptosomes isolated from rat brain cortex

Dielectric measurements were performed on the suspensions of synaptosomes isolated from rat brain cortex. The synaptosomes in buffered salt media showed typical dielectric dispersions caused by the presence of a thin limiting membrane of sufficiently low conductivity. An analysis of the dielectric data revealed that the electric conductivity of the synaptosome interior was about 37 % of the external medium conductivity under isotonic conditions and that the dielectric constant for the interior phase was about 35. The membrane capacitance (0.7 ΜF cm⁻²) remained constant irrespective of nature and concentration of the univalent salts examined. Significant reduction in both the conductivity and the dielectric constant of the internal phase can be explained theoretically provided that some intra-synaptosomal structure (synaptic vesicles and/or small mitochondria) of non-conducting nature occupies about 50 % of the particulate volume, the remainder being in equilibrium with the external salt medium.     

Dielectric properties of synaptosomes isolated from rat brain cortex.

Dielectric measurements were performed on the suspensions of synaptosomes isolated from rat brain cortex. The synaptosomes in buffered salt media showed typical dielectric dispersions caused by the presence of a thin limiting membrane of sufficiently low conductivity. An analysis of the dielectric data revealed that the electric conductivity of the synaptosome interior was about 37% of the external medium conductivity under isotonic conditions and that the dielectric constant for the interior phase was about 35. The membrane capacitance (0.7 muF cm-2) remained constant irrespective of nature and concentration of the univalents salts examined. Significant reduction in both the conductivity and the dielectric constant of the internal phase can be explained theoretically proveded that some intra-synaptosomal structural (synaptic vesicles and/or small mitochondria) of non-conducting nature occupies about 50% of the particulate volume, the remainder being in equilibrium with the external salt medium.     

A new photometric method for oxygen consumption measurements in cell suspensions

A new technique is described for measuring O2 consumption rates and O2 concentrations in suspensions of respiring cells. Aliquots of a cell suspension kept in a special thermostated precision syringe are injected into the measuring system in defined time intervals. The O2 content of these samples is determined photometrically, as reported previously. The O2 consumption per cellular wet weight and/or per single cell can be calculated from the cell volume fraction, the physical density, the cell concentration in the suspension, and the time-dependent decline of the O2 concentration in the precision syringe. The minimum detectable amount of O2 is 0.1 microliter O2, which corresponds to 0.001 (vol/vol) of O2 if a 100-microliters sample of suspended cells is analyzed. Reproducibility of the O2 consumption measurement is 9% of the measured value. The advantages offered by this method are the straightforward calibration in absolute terms, the short time required for one analysis (2-6 min), a high sensitivity, the simultaneous determination of overall O2 concentration and O2 consumption rates in cell suspensions, and the great variability in the application.     

低密度和条件培养对红豆杉细胞生长的影响

红豆杉种胚来源的细胞,在改良Ｂ５液体培养基中继代培养的临界接种密度为鲜重４０ｇ／Ｌ．低密度培养下,１０－１６ｄ的条件培养液（ＣＭ）与新鲜培养液按５７：４３的比例混合时,能显著缩短细胞生长的延迟期,提高生长率,１００Ｌ生物反应器中,按４５．５％体积分数添加条件培养液,在鲜重２７ｇ／Ｌ低接种密度下培养５周,生物量增长９倍,达干重１４．３ｇ／Ｌ．对内源植物激素、精胺、维生素和氨基酸等的比较分析表明,吲哚     

Correlation between biomass and medium conductivity in suspension cultures of rice cells

Summary An intercept in the linear relationship between the biomass increase and decrease in medium conductivity was found in suspension cultures of rice cells. It was due to the. consumption of medium salts by the cells during the lag phase. An equation was established for accurate monitoring of the cell growth which aids understanding of the cell physiology especially during the initial stage of cell growth.     

A nondestructive method for measuring electrical conductivity of intracellular matter of tissue in situ

Summary A nondestructive method has been developed for measuring electrical conductivity of intracellular matter. The method is based on easily measurable electric parameters of cells in suspension. Experiments were made using suspensions of erythrocytes in a physiological salt solution. Results confirmed suitability of the method for determining conductance of the cytoplasm and were in close agreement with those reported by other authors. The method allows the determination of intracellular conductivity in tissue as well.     

A simple method for the measurement of bacterial particle conductivities

A method was developed for the measurement of the bacterial particle conductivity, based on the measurement of the conductivity of a bacterial cell suspension sigma(s) and the suspending medium sigma(m). A line plotted through sigma(s) - sigma(m) versus sigma(m) crosses the x-axis at sigma(m) = sigma(p), independent of the bacterial cell concentration. The method does not require anything more complex than a centrifuge and a conductivity meter. Knowledge of the bacterial particle conductivity is of importance in, for example, the dielectrophoretic separation, manipulation and trapping of bacterial cells, as well as the study of their physiological state.     

Organic matter transport across a macrotidal estuary grandient: the Seine estuary,Fance

The macrotidal Seine estuary receives mineral and organic fluxes from a strongly industrialized basin. Upstream-downstream analysis of POC, polysaccharides and phenolic compounds in the particulate fraction indicated clearly the continental inflow. Within the maximum turbidity zone, the organic matter/suspended particulate matter ratio is lower than elsewhere which results from the dilution effect. Particulate tracers were quantified using grain size analysis and S.E.M. techniques. These sedimentary data defined the ratio of marine to continental particulate flux and mechanisms controlling the suspended particulate matter load within the estuarine mixing zone (resuspension, frontal enrichment and turbulent mixing processes). Organic parameters confirm the data on the mineral matter and were used to distinguish between the marine and continental inputs, from the upstream and downstream samples. Between the two endmembers, variations in organic parameters were controlled mainly by the hydrodynamics in the estuarine zone, rather than by salinity changes.     

Oxygen inhibition of nitrogen fixation in cell-free preparations of blue-green algae

1. Factors inhibiting N₂ fixation in cell-free preparations of two blue-green algae, Anabaena cylindrica and Chlorogloea fritschii were investigated.

2. ¹⁵N uptake by particulate cell-free fractions of A. cylindrica was less in the light than in the dark, indicating some photo-inhibition of N₂ fixation.

3. Anaerobic conditions during preparation and incubation of particulate cell-free fractions of C. fritschii greatly increased their N₂-fixing ability.

4. Soluble cell-free material was found to have an inhibitory effect on N₂ fixation by particulate cell-free preparations obtained from A. cylindrica. This inhibition was approximately proportional to the amount of soluble fraction supplied to the particulate material. Dialysis of the soluble cell-free fraction did not remove the inhibition which has therefore been attributed to the presence of a soluble enzyme, possibly an oxidase.

5. Dialysis of the soluble cell-free fraction or of particulate material suspended with soluble cell-free fraction resulted in a slight decrease of ¹⁵N uptake, indicating the removal by dialysis of small-molecule factors required for N₂ fixation.

6. The presence of sulphydryl agents increased the rate of N₂ fixation in cell-free preparations of A. cylindrica.     

Microbial biomass on particulate organic matter in seawater of the euphotic zone

Microbial biomass on suspended organic matter in seawater of the euphotic zone of Saanich Inlet was investigated. The viable microorganisms were measured by the glucose-uptake method. Microbial carbon on particulate organic matter in seawater was determined to be, on the average, 9.9 μg of C/liter, and there was a regression relationship as y = 0.0062 x − 1.79 with an unbiased variance V_yx^1/2 = 0.38, where x = particulate organic carbon in seawater (micrograms of C/liter) and y = logarithm of microbial carbon (micrograms of C/liter).     

Wave-induced sediment resuspension in the Öre estuary,northern Sweden

Material transported by the Öre River in northern Sweden is all deposited within the estuary which means that resuspension is necessary for the transport of particles out of the estuary. Wave-induced sediment resuspension in the estuary was studied by monitoring the distribution of suspended particles during a resuspension-redeposition cycle. The particle concentration in the water mass was measured with a light scattering probe, calibrated by comparison with the amount of particles collected on a filter.After a long period with calm weather and a low river input less than 100 tonne of suspended particulate matter was present in the estuary. However, during a period with stormy conditions significant resuspension of sediment particles occurred within the estuary. Two days after the storm approximately 1125 tonne of suspended particulate matter was found in the estuary. Most (61%) of the suspended matter was found in the deepest third of the water column, although up to 17% was present in the top third of the water column. The total load of particulate matter in the water column remained constant until day four after the storm, but a significant redistribution of the particulate matter occurred both in the vertical and horizontal directions. Nine days after the storm, a significant amount of particles (c. 350 tonnes) was still in suspension.     

植物叶片吸滞PM2.5等大气颗粒物定量研究方法初探——以毛白杨为例

以毛白杨为例,提出一种利用激光粒度仪和天平定量评估植物叶片吸滞细颗粒物(PM_2.5,直径d≤2.5 μm)等大气颗粒物能力的方法——洗脱称量粒度分析法(EWPA),实现了对植物叶片吸滞大气颗粒物质量和粒径分布的直接、准确测定,可操作性强.首先,进行预试验对试验方法的稳定性进行检验;其次,通过对叶片进行清洗、离心洗液、烘干等步骤收集其吸滞的颗粒物,然后对颗粒物称量,并采用激光粒度仪测定颗粒物的粒径分布;最后,利用叶面积和林分叶面积指数数据换算得到单位面积叶片和林分的各径级颗粒物吸滞量.在北京市奥林匹克森林公园内一片毛白杨林分(27 d未经历降雨)中应用该法,测得毛白杨叶片吸滞大气颗粒物的粒径均值为17.8 μm,吸滞PM_2.5、可吸入颗粒物(PM₁₀,d≤10 μm)和总悬浮颗粒物(TSP,d≤100 μm)的体积百分比分别为13.7%、47.2%和99.9%;叶片的PM_2.5、PM₁₀、TSP和总颗粒物吸滞量分别为8.88×10^-6、30.6×10^-6、64.7×10^-6和64.8×10^-6 g·cm^-2;林分的PM_2.5、PM₁₀、TSP和总颗粒物吸滞量分别为0.963、3.32、7.01和7.02 kg·hm^-2.     

A population balance equation model of aggregation dynamics in Taxus suspension cell cultures

The nature of plant cells to grow as multicellular aggregates in suspension culture has profound effects on bioprocess performance. Recent advances in the measurement of plant cell aggregate size allow for routine process monitoring of this property. We have exploited this capability to develop a conceptual model to describe changes in the aggregate size distribution that are observed over the course of a Taxus cell suspension batch culture. We utilized the population balance equation framework to describe plant cell aggregates as a particulate system, accounting for the relevant phenomenological processes underlying aggregation, such as growth and breakage. We compared model predictions to experimental data to select appropriate kernel functions, and found that larger aggregates had a higher breakage rate, biomass was partitioned asymmetrically following a breakage event, and aggregates grew exponentially. Our model was then validated against several datasets with different initial aggregate size distributions and was able to quantitatively predict changes in total biomass and mean aggregate size, as well as actual size distributions. We proposed a breakage mechanism where a fraction of biomass was lost upon each breakage event, and demonstrated that even though smaller aggregates have been shown to produce more paclitaxel, an optimum breakage rate was predicted for maximum paclitaxel accumulation. We believe this is the first model to use a segregated, corpuscular approach to describe changes in the size distribution of plant cell aggregates, and represents an important first step in the design of rational strategies to control aggregation and optimize process performance.     

样本量对云南松幼苗生物量模型构建及预估精度的影响

为探究不同样本量对生物量模型构建及建模精度的影响，以实际调查的20个家系，共615株云南松幼苗为例，通过编写计算机程序建立进行简单随机抽样，构建不同样本量云南松幼苗各器官及单株生物量异速生长方程。利用决定系数（R²）、估计值标准误（SEE）、均方根误差（RMSE）、总相对误差（RS）及平均误差绝对值（MAB），对模型拟合优度与精度进行比较分析。结果表明：幂函数方程可较好地用于估测云南松幼苗生物量；随着样本量的增加模型精度评估指数MAB呈幂函数形式逐渐减小；当样本数量小于200时MAB较为敏感，模型精度较差，样本量大于200时，其精度随样本量逐渐增加，但变化幅度逐步减小并趋于稳定。因此，根据MAB的变化趋势，样本量达到200时可以构建精度较高且稳定模型。     

CONDUCTIVITY AS A MEASURE OF THE PERMEABILITY OF SUSPENDED CELLS

The problem of determining by means of measurements of electrolytic conductance the permeability of living cells in suspension is considered in some detail and it is pointed out that several factors, usually neglected, have an important influence on the interpretation of such studies. These are: 1. The relative volume and the shape of cells, which are responsive to changes in osmotic pressure and constitution of the surrounding solution. The sources of error in various methods of determining the true volume of red blood cells in a suspension are explained. The hematocrit method appears to be the most reliable method in this case. 2. The proportion of living cells, which is especially to be regarded in the case of suspensions of bacteria. It is shown that this may be very high when appropriate cultural methods are used. The conductance of the dead cells must also be taken into account. 3. The progressive nature of the changes occurring during the course of an experiment. Approximate accuracy may be obtained by proper interpolation. 4. The conductivity of the protoplasm itself, which varies in response to variations is that of the surrounding fluid. It is emphasized that cells, and in particular red blood cells, are not to be regarded as stable non-conducting particles, but rather as labile and as permeable to electrolytes. It is shown that the available data support this interpretation.