Comprehensive manipulation of glycosylation profiles across development scales

The extent and pattern of glycosylation on therapeutic antibodies can influence their circulatory half-life, engagement of effector functions, and immunogenicity, with direct consequences to efficacy and patient safety. Hence, controlling glycosylation patterns is central to any drug development program, yet poses a formidable challenge to the bio-manufacturing industry. Process changes, which can affect glycosylation patterns, range from manufacturing at different scales or sites, to switching production process mode, all the way to using alternative host cell lines. In the emerging space of biosimilars development, often times all of these aspects apply. Gaining a deep understanding of the direction and extent to which glycosylation quality attributes can be modulated is key for efficient fine-tuning of glycan profiles in a stage appropriate manner, but establishment of such platform knowledge is time consuming and resource intensive. Here we report an inexpensive and highly adaptable screening system for comprehensive modulation of glycans on antibodies expressed in CHO cells. We characterize 10 media additives in univariable studies and in combination, using a design of experiments approach to map the design space for tuning glycosylation profile attributes. We introduce a robust workflow that does not require automation, yet enables rapid process optimization. We demonstrate scalability across deep wells, shake flasks, AMBR-15 cell culture system, and 2 L single-use bioreactors. Further, we show that it is broadly applicable to different molecules and host cell lineages. This universal approach permits fine-tuned modulation of glycan product quality, reduces development costs, and enables agile implementation of process changes throughout the product lifecycle.     

用硅藻群集指数(DAIpo)和河流污染指数(RPId)评价珠江广州河段的水质状况

用硅藻计于1990年采集珠江广州河段的9个采样点的附着硅藻群落．共得硅藻24个属,170个分类单位,其中,适腐性种8个,广腐性种84个,仍腐性种78个．群落相似性聚类分析表明,位于前、后航道中部的员村（st3）,新造（st8）,沙浴村（si10）三点的硅藻群落结构是为相似,向上,下游相似性逐渐减小．用硅藻群集指数（DAIpo）和河流污染指数（RPId）对各采样断面的水质进行分析表明：大盛（st12）和莲花山（st5）两点受污染最轻,属β-寡污带,黄沙点（st2）污染最严重,属α-中污带．对照点鸦岗（St1）也受到一定的污染,属α-寡污带,其它点的污染程度在α-寡污带与β-中污带之间．DAIpo值与BODS存在着显著的负相关（r＝0．86）．珠江广州河段的前、后航道的污染程度相差不大（RPId=56．42,56.26）,均属中度污染．     

Establishing a minimum dataset for soil quality assessment based on soil properties and land-use changes

To assess soil quality, a minimum dataset (MDS) of soil properties has to be proposed commonly through calculating the total load of each candidate soil parameter on all of the qualified principal components by use of principal component analysis (PCA) and Norm-value computation. Considering intensive land-use changes, the method introduced in this study on MDS establishment integrates the quantified contributions of land-use type and land-use duration on each soil parameter by using multivariate analysis and mean multiple comparison. In this way, a MDS maximally representing all candidates with minimal loss of the soil quality information contained by those non-MDS soil parameters is established. The MDS proposed can not only well integrate the quantified influence of land-use changes and land-use duration on soil parameters, but is also quite flexible and extendable with the potential to be extrapolated to assess soil quality in other regions. Based on two sets of soil database obtained separately in 1985 and 2004, two MDSs established are compared with each other. It is found that only quite a small change in MDS components occurs during a 20-year period. For a better assessment of soil quality, it seems necessary to examine on what kind of temporal scale and how much MDS will change for a site-specific area with intensive land-use changes.     

Molecular detection of bacterial indicators in cosmetic/pharmaceuticals and raw materials

PCR assays were compared with standard microbiological methods for rapid detection of the United States Pharmacopoeia (USP) bacterial indicators in artificially contaminated samples of raw materials and cosmetic/pharmaceutical products. DNA primers containing the specific sequences of the uidA gene of the β-glucuronidase enzyme for Escherichia coli, the membrane lipoprotein gene oprL for Pseudomonas aeruginosa, and the 16S ribosomal gene for Staphylococcus aureus were used for detection in the PCR reaction. Contaminated samples were incubated for 24 h at 35°C. After incubation in broth media with and without 4% Tween 20, samples were streaked on selective growth media. After 5–6 days, all microbial indicators were morphologically and biochemically identified using standard methods while detection and identification by the PCR-based assays was completed within 27–30 h. Rapid PCR detection of E. coli, S. aureus, and P. aeruginosa will allow a faster quality evaluation and release of raw materials and cosmetic/pharmaceutical products sensitive to microbial contamination. Received 21 June 1998/ Accepted in revised form 11 January 1999     

Foraging strategies and growth inhibition in five daphnids feeding on mixtures of a toxic cyanobacterium and a green alga

1. Laboratory experiments were used to study the feeding, growth and reproduction of five daphnids in mixtures of a toxic cyanobacterium, Microcystis aeruginosa, and a green alga, Scenedesmus acutus. The mixtures included 0%, 20%, 50%, 80% and 100% Microcystis with a total food concentration of 0.5 mg C L^??1 in each treatment. The feeding rate was measured after 1 and 24 h of acclimatization to the mixtures. 2. Toxic Microcystis inhibited feeding in all the species, but they exhibited an unexpected diversity and complexity in the pattern of feeding inhibition. Daphnia magna exhibited the strongest inhibition of feeding after 1 h of exposure to toxic food, but had substantially recovered after 24 h in the same mixtures. This pattern of inhibition and recovery may balance the benefits of reduced ingestion of toxin with the disadvantage of a reduced energy intake. 3. All five daphnids grew quickly in the Scenedesmus control, whereas growth and reproduction declined with an increasing proportion of the toxic alga in the diet. Daphnia pulicaria showed the least inhibition of growth and reproduction, D. pulex showed the strongest inhibition and the three remaining species exhibited intermediate sensitivity. 4. Estimates of gross growth efficiency (GGE; growth/ingestion) provided a means for discriminating between the effects of feeding inhibition and direct toxicity on zooplankton growth. Daphnia pulex exhibited a sharp decline in GGE, suggesting that growth inhibition was a result of both feeding inhibition and direct toxicity. In contrast, D. magna exhibited a nearly constant GGE, indicating that feeding inhibition accounted for its decline in growth. However, two Daphnia species (i.e. D. pulicaria and D. galeata) exhibited improbable increases in GGE with toxic cyanobacteria, suggesting that their feeding rates were underestimated. Growth assays with sensitive and resistant zooplankton species are proposed for testing the potential impacts toxic cyanobacteria in lakes.     

Microbiological and chemical quality of a traditional salted-fermented fish (Hout-Kasef) product of Jazan Region,Saudi Arabia

The Hout-Kasef is traditional salted fermented fish product of natural fermentation of salted mullet fish of coastal area of Jazan region of Saudi Arabia. The present study was carried out to investigate the microbiological and chemical characteristic of Hout-Kasef. A total of twenty-four salted fish samples were purchased from fish market in Jazan and Abu-Arish at different times of the year. The microbial studies of salted-fermented fish revealed a total bacterial count ranging from 2.81 to 4.72 Log₁₀ CFU/g, yeast and mold counts ranging from 0.48 to 3.14 Log₁₀ CFU/g, total staphylococci count 2.71–3.85 Log₁₀ CFU/g, halophile bacteria count 3.26–5.14 Log₁₀ CFU/g, and coliforms count <1 Log₁₀ CFU/g. However, pathogenic bacteria such as Listeria monocytogenes, Vibrio spp., Campylobacter spp. and Yersinia species were not detected. The major bacteria species isolated and identified from the salted fermented fish were Bacillus Subtilus, Bacillus mycoides, Bacillus licheniformis, Bacillus pumilus, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus hominis, Staphylococcus xylosus, Staphylococcus saprophticus and Staphylococcus cahnii subsp cahnii. The chemical analysis of salted fermented fish showed high content of moisture (47.96%), protein (25.71%), ash (19.6%) and salt (15.19%) but low contents of lipid (7.25%). The salted-fermented fish also showed high level of total volatile basic nitrogen (78.86 mg/100 gm sample) and thiobarbutric acid number (32.32 mg malonaldehyde/kg) with a pH value of pH 6.3. Finally, this study showed the presence of gram positive and gram negative bacteria in the fish product. The predominant microorganisms found were Bacillus and Staphylococcus spp. The fish product had high content of salt and TVB-N levels.     

新发缺血性脑卒中患者实施单病种质量管理的预后评价

目的 调查分析上海市某三甲医院缺血性脑卒中单病种质量管理实施情况并评价其效果。方法 采用回顾性调查的方法,以NIHSS评分为疗效评价依据,对新发缺血性脑卒中单病种质量管理实施组和未实施组的预后情况进行分析评估。结果 在2组基线资料均衡的情况下,缺血性脑卒中单病种质量管理实施组的疗效显著优于未实施组,尤其是发病初始疾病严重程度一般者的疗效最好。结论 缺血性脑卒中单病种质量管理通过对诊疗过程的质量控制,规范了诊疗行为,改善了新发缺血性脑卒中患者的预后。     

Prey preference and egg production of the carabid beetleAgonum dorsale

In order to elucidate whether the aphidRhopalosiphum padi is low quality food for the carabid beetleAgonum dorsale, as it has previously been found to be for cereal spiders, we performed a series of experiments using fruit fliesDrosophila melanogaster as a standard alternative prey (‘control prey’): 1) Prey preference, 2) Aphid consumption for satiated and starved beetles, 3) Egg production on different diets and 4) Preference experiments with fruit flies coated with a taste of different prey types. Other alternative prey types used were earthworms and sciarid midges, all potential prey in the field.A. dorsale has a low preference for aphids compared to fruit flies. Apparently there is an upper limit to the consumption of aphids independent of hunger and much below the beetles' total food demand. Egg production on a pure diet of aphids is lower than on a pure diet of fruit flies; it is extremely low on a pure diet of earthworms; the highest fecundity is found on a mixed diet. As fruit flies coated with a taste of aphids or sciarid midges are less prefered than flies covered with a taste of fruit flies there may be a chemical factor, acting through taste, involved in determining prey preference.     

THE EFFECTIVE SIZE OF A HIERARCHICALLY STRUCTURED POPULATION

A knowledge of the effective size of a population (N_e) is important in understanding its current and future evolutionary potential. Unfortunately, the effective size of a hierarchically structured population is not, in general, equal to the sum of its parts. In particular, the inbreeding structure has a major influence on N_e. Here I link N_e to Wright's hierarchical measures of inbreeding, F_IS and F_ST, for an island-structured population (or metapopulation) of size N_T. The influence of F_ST depends strongly on the degree to which island productivity is regulated. In the absence of local regulation (the interdemic model), interdemic genetic drift reduces N_e. When such drift is combined with local inbreeding under otherwise ideal conditions, the effects of F_IS and F_ST are identical: increasing inbreeding either within or between islands reduces N_e, with N_e = N_T/[(1 + F_IS)(1 + F_ST) ? 2F_ISF_ST]. However, if islands are all equally productive because of local density regulation (the traditional island model), then N_e = N_T/[(1 + F_IS)(1 –F_ST)] and the effect of F_ST is reversed. Under the interdemic model, random variation in the habitat quality (and hence productivity) of islands act to markedly decrease N_e. This variation has no effect under the island model because, by definition, all islands are equally productive. Even when no permanent island structure exists, spatial differences in habitat quality can significantly increase the overall variance in reproductive success of both males and females and hence lower N_e. Each of these basic results holds when other nonideal factors are added to the model. These factors, deviations from a 1:1 sex ratio, greater than Poisson variance in female reproductive success, and variation in male mating success due to polygynous mating systems, all act to lower N_e. The effects of male and female variance on N_e have important differences because only females affect island productivity. Finally, it is noted that to use these relationships, F_IS and F_ST must be estimated according to Wright's definition (and corrected to have a zero expectation under the null model). A commonly used partitioning (θ, θ_g) can be biased if either island size or the number of islands is small.     

中国生物多样性在线数据处理平台的构建

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