Characterization of TAP Ambr 250 disposable bioreactors,as a reliable scale‐down model for biologics process development

Demands for development of biological therapies is rapidly increasing, as is the drive to reduce time to patient. In order to speed up development, the disposable Automated Microscale Bioreactor (Ambr 250) system is increasingly gaining interest due to its advantages, including highly automated control, high throughput capacity, and short turnaround time. Traditional early stage upstream process development conducted in 2 ‐ 5 L bench‐top bioreactors requires high foot‐print, and running cost. The establishment of the Ambr 250 as a scale‐down model leads to many benefits in process development. In this study, a comprehensive characterization of mass transfer coefficient (k_La) in the Ambr 250 was conducted to define optimal operational conditions. Scale‐down approaches, including dimensionless volumetric flow rate (vvm), power per unit volume (P/V) and k_La have been evaluated using different cell lines. This study demonstrates that the Ambr 250 generated comparable profiles of cell growth and protein production, as seen at 5‐L and 1000‐L bioreactor scales, when using k_La as a scale‐down parameter. In addition to mimicking processes at large scales, the suitability of the Ambr 250 as a tool for clone selection, which is traditionally conducted in bench‐top bioreactors, was investigated. Data show that cell growth, productivity, metabolite profiles, and product qualities of material generated using the Ambr 250 were comparable to those from 5‐L bioreactors. Therefore, Ambr 250 can be used for clone selection and process development as a replacement for traditional bench‐top bioreactors minimizing resource utilization during the early stages of development in the biopharmaceutical industry. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:478–489, 2017     

基于主因子分析的生态社区评价方法研究——以上海外环以内区域为例

从生态系统和可持续发展理论出发,构建了生态社区评价的指标体系,运用主因子分析方法建立了生态社区评价模型;并在遥感与GIS技术支持下,对上海中心城区做了生态社区评价研究.结果表明,在空间格局上,社区生态水平差异明显,无论是在社区内部还是社区之间,各主因子分布极不平衡;总体看来,浦西核心区的大部分社区,其规模结构比较合理,服务设施比较完善,然而部分社区的生态环境质量急需要改善,部分社区的居住条件也需要进一步完善;而浦东一些社区,尽管生态指数较高,生态环境质量较好,但由于基础配套设施不够健全,规模结构不够均衡,道路交通系统尚待进一步完善.     

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.     

Predator and detritivore niche width helps to explain biocomplexity of experimental detritus-based food webs in four aquatic and terrestrial ecosystems

In the study of food webs, the existence and explanation of recurring patterns, such as the scale invariance of linkage density, predator–prey ratios and mean chain length, constitute long-standing issues. Our study focused on litter-associated food webs and explored the influence of detritivore and predator niche width (as δ¹³C range) on web topological structure. To compare patterns within and between aquatic and terrestrial ecosystems and take account of intra-habitat variability, we constructed 42 macroinvertebrate patch-scale webs in four different habitats (lake, lagoon, beech forest and cornfield), using an experimental approach with litterbags. The results suggest that although web differences exist between ecosystems, patterns are more similar within than between aquatic and terrestrial web types. In accordance with optimal foraging theory, we found that the niche width of predators and prey increased with the number of predators and prey taxa as a proportion of total taxa in the community. The tendency was more marked in terrestrial ecosystems and can be explained by a lower per capita food level than in aquatic ecosystems, particularly evident for predators. In accordance with these results, the number of links increased with the number of species but with a significantly sharper regression slope for terrestrial ecosystems. As a consequence, linkage density, which was found to be directly correlated to niche width, increased with the total number of species in terrestrial webs, whereas it did not change significantly in aquatic ones, where connectance scaled negatively with the total number of species. In both types of ecosystem, web robustness to rare species removal increased with connectance and the niche width of predators. In conclusion, although limited to litter-associated macroinvertebrate assemblages, this study highlights structural differences and similarities between aquatic and terrestrial detrital webs, providing field evidence of the central role of niche width in determining the structure of detritus-based food webs and posing foraging optimisation constraints as a general mechanistic explanation of food web complexity differences within and between ecosystem types.     

孝感城区中老年对慢性病及社区管理现状认知情况的调查研究

目的:调查孝感城区中老年对慢性病及其社区管理现状认知情况。方法:选择2010年7月至2015年7月在孝感城区3个社区的500名中老年人作为研究对象,利用自拟调查问卷对所有受试者进行数据调查,分析孝感城区中老年对慢性病的认知情况,及对慢性病社区管理现状的认知情况,分析影响慢性病认知的因素。结果:在慢性病种类中,孝感城区中老年人对于高血压和糖尿病,及风湿性关节炎与慢性支气管炎的认知率较高,而对于癌症、偏瘫及血栓的认知率较低,差异有统计学意义(P0.05)。在慢性病的危险因素中,孝感城区中老年人对于缺乏锻炼、吸烟及膳食不合理的认知率明显较高,而对于酗酒、缺乏保健知识及悲观情绪的认知率明显较低,差异有统计学意义(P0.05)。在慢性病社区管理现状上,孝感城区中老年对于指导其改变生活习惯的认知率较高,为62.00%,而在进行三级预防、强化慢性病自我管理、定期举办慢性病专题讲座以及建立社区卫生的信息服务平台等方面的认知率明显较低,分别为22.40%、36.80%、42.20%及21.80%,差异有统计学意义(P0.05)。Logistic分析显示,文化程度低、经济收入低、未患慢性病及年龄≥60岁均为慢性病认知的危险因素。结论:孝感城区中老年对慢性病及其社区管理现状认知仍存在一定的不足,相关社区卫生管理部门应针对问题的影响因素,积极采取措施,改善社区管理现状。     

Source-sink landscape theory and its ecological significance

Exploring the relatiouships between landscape pattern and ecological processes is the key topic of landscape ecology,for which,a large number of indices as well as landscape pattern analysis model were developed.However,one problem faced by landscape ecologists is that it is hard to link the landscape indices with a specific ecological process.Linking landscape pattern and ecological processes has become a challenge for landscape ecologists."Source" and "sink" are common concepts used in air pollution research,by which the movement direction and pattern of different pollutants in air can be clearly identified.In fact,for any ecological process,the research can be considered as a balance between the source and the sink in space.Thus,the concepts of "source" and "sink" could be implemented to the research of landscape pattern and ecological processes.In this paper,a theory of sourcesink landscape was proposed,which include:(1) In the research of landscape pattern and ecological process,all landscape types can be divided into two groups,"source"landscape and "sink" landscape."Source" landscape contributes positively to the ecological process,while "sink" landscape is unhelpful to the ecological process.(2) Both landscapes are recognized with regard to the specific ecological process."Source" landscape in a target ecological process may change into a "sink"landscape as in another ecological process.Therefore,the ecological process should be determined before "source"or "sink" landscape were defined.(3) The key point to distinguish "source" landscape from "sink" landscape is to quantify the effect of landscape on ecological process.The positive effect is made by "source" landscape,and the negative effect by "sink" landscape.(4) For the same ecological process,the contribution of "source" landscapes may vary,and it is the same to the "sink"landscapes.It is required to determine the weight of each landscape type on ecological processes.(5) The sourcesink principle can be applied to non-point source pollution control,biologic diversity protection,urban heat island effect mitigation,etc.However,the landscape evaluation models need to be calibrated respectively,because different ecological processes correspond with different source-sink landscapes and evaluation models for the different study areas.This theory is helpful to further study landscape pattern and ecological process,and offers a basis for new landscape index design.     

Modeling Neisseria meningitidis B metabolism at different specific growth rates

Neisseria meningitidis is a human pathogen that can infect diverse sites within the human host. The major diseases caused by N. meningitidis are responsible for death and disability, especially in young infants. At the Netherlands Vaccine Institute (NVI) a vaccine against serogroup B organisms is currently being developed. This study describes the influence of the growth rate of N. meningitidis on its macro-molecular composition and its metabolic activity and was determined in chemostat cultures. In the applied range of growth rates, no significant changes in RNA content and protein content with growth rate were observed in N. meningitidis. The DNA content in N. meningitidis was somewhat higher at the highest applied growth rate. The phospholipid and lipopolysaccharide content in N. meningitidis changed with growth rate but no specific trends were observed. The cellular fatty acid composition and the amino acid composition did not change significantly with growth rate. Additionally, it was found that the PorA content in outer membrane vesicles was significantly lower at the highest growth rate. The metabolic fluxes at various growth rates were calculated using flux balance analysis. Errors in fluxes were calculated using Monte Carlo Simulation and the reliability of the calculated flux distribution could be indicated, which has not been reported for this type of analysis. The yield of biomass on substrate (Y(x/s)) and the maintenance coefficient (m(s)) were determined as 0.44 (+/-0.04) g g(-1) and 0.04 (+/-0.02) g g(-1) h(-1), respectively. The growth associated energy requirement (Y(x/ATP)) and the non-growth associated ATP requirement for maintenance (m(ATP)) were estimated as 0.13 (+/-0.04) mol mol(-1) and 0.43 (+/-0.14) mol mol(-1) h(-1), respectively. It was found that the split ratio between the Entner-Doudoroff and the pentose phosphate pathway, the sole glucose utilizing pathways in N. meningitidis, had a minor effect on ATP formation rate but a major effect on the fluxes going through for instance the citric-acid cycle. For this reason, we presented flux ranges for underdetermined parts of metabolic network rather than presenting single flux values, which is more commonly done in literature.     

Application of DNA amplification fingerprinting (DAF) to mixed culture bioreactors

Summary The use of DNA amplification fingerprinting (DAF) as a tool for monitoring mixed microbial populations in bioreactors was evaluated. Short (8-mer or 10-mer) oligonucleotides were used to prime DNA extracts from various biological reactors during polymerase chain reaction (PCR) amplification. The reactors examined in this study included two sets of anaerobic stirred tank continuous flow bioreactors. One set of anaerobic reactors was operated under methanogenic conditions and one set was operated under sulfate-reducing conditions. The anaerobic reactor communities in the methanol-fed reactors showed extensive DAF homology. DAF was also applied to a fixed-film azo dye degrading reactor to examine the degree of uniformity of colonization of the substratum in representative regions of the reactor. This method is a quick and relatively inexpensive means of monitoring microbial community structure during biological processes. Since no cultivation of the sample is involved, the genetic profile of the community is not biased by outgrowth conditions. DAF profiles may be useful for comparisons of population changes over time or of bench-scale vs pilot-scale reactors but not adequate for assessing community diversity.     

Impact of superheated steam roasting on changes in antioxidant and microstructure properties of raw and processed cocoa cotyledon

This research focused on the roasting of cocoa beans at 184 °C for 16 min duration in a superheated steam oven using two separate modes of heating: convection mode and superheated steam mode. After roasting, the antioxidant properties of the cooked cocoa were assessed as ferric reducing antioxidant power activity (FRAP), DPPH radical scavenging activity, total flavonoid content (TFC) and total phenol content (TPC). The micro structural properties of raw and processed cocoa beans were observed using scanning electron microscopy (SEM). As discovered in the scan, conventional roasting showed a nearly complete rapture of the cytoplasmic network system and the destruction of the organelles, whereas superheated steam mode showed satisfactory images. Studies indicated that superheated steam roasting preserved significantly (p < 0.05) greater antioxidant properties as opposed to conventional method of roasting.