Comparison of optimization methods for fed-batch cultures of hybridoma cells

Two methods for the calculation of optimal trajectories for the input variables of a fed-batch culture of hybridoma cells are compared. It pointed out that a gradient method based on Pontryagins' minimum principle based yields a significant better performance with respect to computational effort and the calculated minimum than a dynamic programming approach which has been presented in a previous paper [1] as the most suitable method.     

Human beta-glucuronidase. Studies on the effects of pH and bile acids in regard to its role in the pathogenesis of cholelithiasis

Human bile contains a considerable amount of endogenous beta-glucuronidase. The effects of pH and bile acids on its activity have been studied in regard to its role in the pathogenesis of cholelithiasis. beta-Glucuronidase, purified from human liver to homogeneity, was structurally stable between pH 4 and 10, but was active only over a much narrower range of pH, with a pH optimum of 5.2. The inactivation below pH 4 was due to its irreversible denaturation, whereas the inactivation at higher pH was due to a true reversible pH effect on the enzyme velocity. Kinetic studies revealed that hydrogen ion acted as a substrate-directed activator of the free enzyme, but not the enzyme-substrate complex, with a molecular dissociation constant of 4 X 10(-6). The enzyme activity was not affected by unconjugated bile acids, primarily due to their extremely low water solubility. Conjugated bile acids, on the other hand, exerted heterogeneous and pH-dependent effects on the enzyme. At pH 5.2, taurocholic acid and glycocholic acid were substrate-directed activators of the enzyme; taurochenodeoxycholic acid and taurodeoxycholic acid, competitive inhibitors; and glycochenodeoxycholic acid and glycodeoxycholic acid, mixed inhibitors. At pH 7.0 all taurine and glycine conjugates behaved as substrate-directed activators. Though beta-glucuronidase activity at pH 7 was only 23% of its maximal activity at pH 5.2, conjugated bile acids tended to restore its activity to a certain extent at pH 7. Thus, endogenous beta-glucuronidase could play a significant role in pigment cholelithiasis.     

Cloning and expression of the Erwinia chrysanthemi asparaginase gene in Escherichia coli and Erwinia carotovora

A genomic library of Erwinia chrysanthemi DNA was constructed in bacteriophage lambda 1059 and recombinants expressing Er. chrysanthemi asparaginase detected using purified anti-asparaginase IgG. The gene was subcloned on a 4.7 kb EcoRI DNA restriction fragment into pUC9 to generate the recombinant plasmid pASN30. The position and orientation of the asparaginase structural gene was determined by subcloning. The enzyme was produced at high levels in Escherichia coli (5% of soluble protein) and was shown to be exported to the periplasmic space. Purified asparaginase from E. coli cells carrying pASN30 was indistinguishable from the Erwinia enzyme on the basis of specific activity [660-700 units (mg protein)-1], pI value (8.5), and subunit molecular weight (32 X 10(3]. Expression of the cloned gene was subject to glucose repression in E. coli but was not significantly repressed by glycerol. Recombinant plasmids, containing the asparaginase gene, when introduced into Erwinia carotovora, caused increased synthesis of the enzyme (2-4 fold higher than the current production strain).     

Predator detection and avoidance by starlings under differing scenarios of predation risk

Practically all animals must find food while avoiding predators.An individual's perception of predation risk may depend on manyfactors, such as distance to refuge and group size, but it isunclear whether individuals respond to different factors ina similar manner. We tested whether flocks of foraging starlingsresponded in the same way to an increased perception of predationrisk by assessing three factors: (1) neighbor distances, (2)habitat obstruction, and (3) recent exposure to a predator.We found that in all three scenarios of increased risk, starlingsreduced their interscan intervals (food-searching bouts), whichincreased the frequency of their vigilance periods. We thenexamined how one of these factors, habitat obstruction, affectedescape speed by simulating an attack with a model predator.Starlings were slower to respond in visually obstructed habitats(long grass swards) and slower when they had their head downin obstructed habitats than when they had their head down inopen habitats. In addition, reaction times were quicker whenstarlings could employ their peripheral fields of vision. Ourresults demonstrate that different sources of increased riskcan generate similar behavioral responses within a species.The degree of visibility in the physical and social environmentaffects both the actual and perceived risk of predation.     

Distribution of the antimonate precipitate in normal and segregated nucleoli after potassium pyroantimonate fixation

Fixation with a saturated aqueous solution of potassium pyroantimonate produces electron-opaque antimonate deposits in the nucleoli of lutheinic and hepatic cells. The comparative analysis between normal and actinomycin D segregated nucleoli fixed either with glutaraldehyde-osmium or potassium pyroantimonate facilitates location of the ring-shaped precipitates in the fibrillar centers and the fine dense ones in the pars fibrosa.     

N-糖蛋白去糖基化酶（PNGase）的研究进展

N-糖蛋白去糖基化酶（PNGase）是一种广泛存在于真菌、植物、哺乳动物中的去糖基化酶，可以水解N-糖蛋白或 N-糖肽上天冬酰胺与寡糖链连接的化学键，并释放出完整的N-寡糖。PNGase在生物体内参与蛋白质降解、器官发育、个体生长等过程。人PNGase基因功能缺陷会导致先天性去糖基化障碍，小鼠PNGase缺陷会导致胚胎致死性，线虫PNGase缺陷使其寿命下降。本文对PNGase在不同物种的分布、蛋白质结构、酶学功能及生物学功能进行阐述，为PNGase的生理病理功能及致病机制的基础研究提供思路，为PNGase作为糖生物学工具酶或药物开发的创新应用研究奠定基础。