The culture of rat myeloma and rat hybridoma cells in a protein-free medium

Y0 is a rat x rat hybridoma cell line, which does not secrete immunoglobulin, produced using a fusion partner derived from the Y3 (Y3,Ag.1.2.3) rat myoloma cell line. Y0 and Y3 have both been widely used as fusion partners in the production of rat x rat hybridomas. Y0 has also been used in recombinant gene technology. Y0 cells grown in shake flask culture, using RPMI 1640 medium with 4mM l-glutamine and 5% foetal bovine serum, reached a maximal cell density of 1.5×10⁶ cells ml^–1 with 86% viability. Y0 cells which has been adapted to grow in ABC protein-free medium reached a maximal density, in shake flask culture, of 8.75×10⁵ cells ml^–1 with 79% viability. An improved protein-free medium, designated W38 medium, was developed. In shake flask culture, W38 medium supported Y0 cell growth to a density of 2.02×10⁶ cells ml^–1 with 96% viability. Two Y3 hybridomas, YID 13.9.4 cells and SAM 618 cells were adapted to growth in W38 medium. For both hybridomas, cell growth and product yield in shake flask culture using W38 medium was superior to that obtained with serum-containing RPMI 1640 medium.Abbreviations F12 Hams F12 medium - DMEM Dulbeccos medium - RPMI RPMI 1640 medium - FBS foetal bovine serum     

Stabile production of a thrombin resistant pro-urokinase derivative (PRO-UKS1) by Namalwa KJM-1 cells adapted to serum-free medium

Pro-UKS1 was designed as a thrombin-resistant derivative of pro-urokinase (pro-UK) by introducing a glycosylation site using site-directed mutagenesis. An expression plasmid for pro-UKS1, pMo1UKS1SEd1-5, was constructed and introduced into Namalwa KJM-1 cells (Hosoiet al., 1988), and cells resistant to G418 and Methotrexate (MTX) were obtained. Amongst them, the highest pro-UKS1 producer (resistant to 500 nM of MTX), clone 41-8, was selected and further characterized. Clone 41-8 was cultured in serum-free ITPSGF medium (Hosoiet al., 1988). Under the conventional conditions, the concentration of pro-UKS1 reached 26 g ml^–1. Addition of glucose and tri-iodothyronine (T₃) improved productivity, and the maximal productivity of pro-UKS1 was 67 g ml^–1 day^–1. In this conditioned medium, content of pro-UKS1 was above 80% of total proteins.Abbreviations BSA bovine serum albumin - dhfr dihydrofolate reductase - HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid - kb kilobase pairs - kDa kilodaltons - MTX Methotrexate - PBS phosphate buffered saline - pro-UK pro-urokinase - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis - T₃ tri-iodothyronine - Tween-PBS phosphate buffered saline containing 0.05% Tween 80     

Induction of autonomous endosperm in Lupinus luteus,Helleborus niger and Melandrium album by in vitro culture of unpollinated ovaries

Autonomous division of the endosperm was induced by in vitro culture of unpollinated ovaries or placenta-attached ovules in Helleborus niger, Lupinus luteus and Melandrium album. The induction frequencies for the three species were 50%, 10–20% and 0.1%, respectively. The endosperms contained up to 20 free nuclei; only a few ovules with 80–420 endosperm nuclei were found. Induction of autonomous division of the endosperm, which is unusual in amphimictic plants, was observed in three new species. No embryos appeared in the ovules. This suggests a developmental independence of the endosperm from the embryo in the culture of unpollinated ovaries or ovules.     

Plant diversity in late medieval cornfields of northern Switzerland

Weed assemblages from late medieval cornfields have been studied for the first time in northern Switzerland. Eleven samples from at least two different grain stores were investigated. The samples were collected from the carbonised remains of six wooden houses built in the late 13th century A.D. and which burnt down in the middle of the 15th century. The weed floras found in the spelt (Triticum spelta) and oats (Avena sativa) indicate a high botanical diversity in the cornfields at harvest time. Although oats are normally a summer crop and spelt a winter crop, both summer and winter crop weeds (as well as many different present-day grassland taxa) were found in each type of grain. Most of the weeds found were perennial plants, which was interpreted as an indication of both extensive tillage of the arable land and application of the three-field rotation system (Dreifelderwirtschaft). The spectra of the two palaeophytocoenoses (assemblages of ancient plant remains) studied suggest that the phytosociological method may not be reliable for classification of the late medieval remains into summer and winter crop weed communities. These findings should provide a better understanding of the development of anthropogenic plant communities, and in particular, the development of crop weed communities.     

Mutations in neuromusculin,a gene encoding a cell adhesion molecule,cause nervous system defects

The Drosophila neuromusculin (nrm) gene encodes an immunoglobulin-like (Ig-like) cell adhesion molecule expressed in the precursors of the embryonic peripheral nervous system (PNS), in the midline precursors of the central nervous system (CNS), and in muscles. During the initial phases of CNS axonogenesis, nrm is expressed in cells involved in the development of commissures and longitudinal tracts. Mutations which alter expression of nrm mRNAs cause aberrant development of commissures and longitudinal axon pathways. Defects in the PNS and muscles of nrm mutants are also observed. In most nrm embryos, abnormal development can be detected in a subset of abdominal segments; however, in approximately 1 of 10 nrm embryos, the defects extend to all segments. Herein, we present evidence that nrm plays an important role in early morphogenesis, possibly by mediating or facilitating inductive cell contacts and movements.     

Recent advances in plant cell cultures in bioreactors

Two key issues in the application of plant-cell-culture technology to the production of valuable secondary metabolites are reviewed: the selection of cell lines with suitable genetic, biochemical and physiological characteristics; and the optimization of bioreactor environments. Although great progress has been made in recent years in the design, selection and optimization of bioreactor hardware, optimization of environmental factors such as medium components, light irradiation and O₂ supply needs detailed investigations for each case. With a better understanding of plant cell metabolism and physiology, further developments in cultivation processes, such as process integration and on-line monitoring and control, can be expected in the near future.J.-J. Zhong and J.-T. Yu are with the Research Institute of Biochemical Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, China T. Yoshida is with the International Center of Cooperative Research in Biotechnology (ICBiotech), Faculty of Engineering, Osaka University, Suita, Osaka 565, Japan.     

Multiple paternity in the red-eyed treefrog Agalychnis callidryas (Cope)

DNA fingerprinting of tadpoles from two different ‘one female-two male’ matings of the red-eyed treefrog, Agalychnis callidryas, revealed multiple paternity of offspring. Offspring were assigned paternity based on bands shared with the putative father, but not shared between putative fathers or with the mother. Paternity was split 44/56% and 36/64% in two matings. These results do not support a hypothesis of sperm priority in access to unfertilized eggs by primary males. Multiple paternity may be commonplace in species of anurans with matings by multiple males.     

苯并［a］芘在土壤－水－水稻系统中传输动力学研究

建立了ＢａＰ在土壤水稻系统中传输模型,并对其在系统各组分中的含量变化做了定量分析。实验求得了ＢａＰ在系统各相间传输速度常数和分配系数。揭示了水中ＢａＰ向水稻体传输路径和它们的速度大小。水稻从水中吸收ＢａＰ的速度比其从土壤中吸收的速度高４倍,水中ＢａＰ向土壤沉积速度比水稻从水中吸收ＢａＰ的速度高７倍。水中ＢａＰ在迁移到达水稻体之前,绝大部分被土壤牢牢吸附而很难被水稻吸收。     

甘蔗抗逆细胞系选择及其生化特性的研究

利用脯氨酸类似物羟脯氨酸(Hyp)的生长抑制作用,筛选出抗Hyp的甘蔗(Saccha-rum sinensis Roxb)细胞变异系R932。抗性系体内游离脯氨酸含量超常积累(×3．2)。而且其体内脯氨酸合成途径中的关键酶γ－谷氨酸激酶对脯氨酸的反馈抑制较不敏感。R932抗PEG和低温能力较供体加强。实验结果表明γ－谷氨酸激酶特性的变化可能导致细胞内脯氨酸的过量积累,脯氨酸的过量积累有利于植物细胞对抗恶劣环境。     

鳜鱼的耗氧率及其池塘养殖

本文较详细地报道了鳜鱼的耗氧率和窒息点,并对鳜鱼和作其饲料的鱼同池饲养的可能性进行了探讨。结果表明,鳜鱼耗氧量和体重正相关（ｒ＝０．９９）,耗氧率与体重反相关（ｒ＝－０．９７）;在水温２０℃,鱼种耗氧率约为０．１４ｍｇ／ｇ．ｈ．,成鱼约为０．１２ｍｇ／ｇ．ｈ．;耗氧量和耗氧率均与水温正相关（ｒ＝０．９０,ｒ＝０．９４）,水温１３—３０℃时,体重２３０±１１．７ｇ的鳜鱼。耗氧量为１４．３１—４２．１３ｍｇ／尾。ｈ．,耗氧率为０．０５９—０．１７５ｍｇ／ｇ．ｈ．;鳜鱼耗氧率昼夜变化与家鱼相反,黄昏至凌晨是高峰期,为０．１２—０．１５ｍｇ／ｇ．ｈ．（Ｔ＝２０℃）,白天是低谷期,为０．０７—０．１０ｍｇ／ｇ．ｈ;;鳜鱼的窒息点与家鱼类较接近,变化范围为０．４５—０．７６ｍｇ／Ｌ;鳜鱼和作其饲料的鱼在同一池塘中饲养,既可持续不断地提供鳜鱼充足的适口饵料,又简单易行,成本低,效益高,有较大的价值。