Studies of cell pellets: I. Electrical properties and porosity.

Cell pellets formed by centrifugation provided a good system to study the osmotic behavior, electroporation, and interaction between cells. Rabbit erythrocyte pellets were used in this study because they were simpler than nucleated cells to model analytically. Structurally, cell pellets possessed properties of porous solid bodies and gels. Electrically, cell pellets were shown to behave as a parallel set of resistance, Rp, and capacitance, Cp. Information on pellet structures was obtained from electric measurements. The pellet resistance reflected the intercellular conductivity (porosity and gap conductivity), whereas the pellet capacitance depended mostly on membrane capacitance. The pellet resistance was more sensitive to experimental conditions. The intercellular gap distance can be derived from pellet porosity measurements, providing the cell volume and surface area were known. Rp increased and relaxed exponentially with time when centrifugation started and stopped; the cycles were reversible. When supernatants were exchanged with solutions containing hypotonic electrolytes or macromolecules (such as PEG) after the pellets were formed, complicated responses to different colloidal osmotic effects were observed. A transient decrease followed by a large increase of Rp was observed after the application of a porating electric pulse, as expected from a momentary membrane breakdown, followed by a limited colloidal-osmotic swelling of pelleted cells. The equilibrium values of Rp, Cp, pellet porosity, and intercellular distances were measured and calculated as functions of cell number, centrifugation force, and ionic strength of the exchanged supernatant. Thus, the structure and properties of cell pellets can be completely characterized by electrical measurements.     

Electrically induced fusion of mammalian cells in the presence of polyethylene glycol

Chinese Hamster Ovary (CHO) cells were fused by subjecting cell suspensions to an exponentially decaying electric pulse in the presence of polyethylene glycol (PEG), Dextran or Ficoll. PEG (MW 1,000, 3,350, 8,000, 10,000 and 18,500), Dextran (MW 71,200) and Ficoll (MW 400,000) were added to the pulsing medium. A single exponential electric pulse with peak field strength of 4 kV/cm, and a half-time of 0.72 msec was used. The combination of two techniques, PEG-induced fusion and electrofusion, resulted in highly efficient fusion of CHO cells. Fusion yields (FY) at different concentrations of these polymers were measured using phase-contrast microscopy. FY was highly dependent on the concentration of PEG in media, while the presence of Dextran and Ficoll had no influence on fusion yield. PEG with MW 8,000 was found to be the most effective in causing cell aggregation, and to give the highest FY (40%). An optimal concentration for fusion was found for PEG of each molecular weight. Diluting cells suspended in higher concentrations of PEG to these optimal concentrations after the pulse application regained the optimal FY. It was concluded that PEG-induced prepulse aggregation and moderate cell swelling immediately after the pulse were important factors in achieving high fusion yields.This work is supported by a grant GM-30969 from the National Institutes of Health. Traveling fellowship to N.G.S. was supported from Foundation Cyrill and Methodius and grant N-189 from MCES of Bulgaria.     

Effect of heavy metals on the methanogenic activity of starch-degrading granules

Summary Heavy metals in electroplating effluent inhibited specific methanogenic activity (SMA) of anaerobic starch-degrading granules. The SMA of granules on the degradation of starch were reduced by 50% when each gram of biomass was in contact individually with 105 mg of zinc, 120 mg of nickel, 180 mg of copper, 310 mg of chromium, or >400 mg of cadmium. Granules had higher toxicity-resistance than flocculent sludge, due to their layered structure.     

人工饲养蒙新河狸的发情交配行为

人工饲养的蒙新河猪其动情周期可划分为４个阶段：动情前期、动情期（发情交配期）、动情后期和停滞期（动情间隔期）。动情期间雌雄河狸均表现出活跃好动,浮躁不安,食量减小;发情交配期雌雄河狸均发出低沉嘶哑的叫声,雄性叫声频率比性略高,爬跨交配是动情期发情的高峰,动情周期各阶段的行为特征,在雌性与阴道涂片细胞学及外生殖器形态、色泽、阴道分泌物周期性变化特征相吻合,在雄性则与睾丸及阴茎形态特征吻合,在人工饲养     

旋毛虫肌幼虫ES抗原的基因克隆及高效表达

作者对编码旋毛虫肌幼虫ES抗原的部分结构基因进行了克隆、鉴定和表达。用RNA PCR技术直接从旋毛虫肌幼虫总RNA中反转录并扩增出0．7kh的靶DNA,酶切分析后将其克隆到融合表达载体pEx3lC中。SDS—PAGE电泳表明,含重组子的大肠杆菌能够表达出一分子量为37kDa的融合蛋白(P37),后者占菌体总蛋白的22%以上,并以包含体形式存在于菌体中。经对纯化后表达蛋白的ELlSA检测,证明它能被猪旋毛虫病阳性血清和抗旋毛虫单克隆抗体识别。研究结果揭示,重组蛋白P37对于研制旋毛虫病诊断抗原和免疫抗原具有潜在的应用价值。     

大鼠心脏血压超负荷诱导左心室HSP70基因表达

本工作应用热休克蛋白７０（ＨＳＰ７０）核酸分子杂交方法,测定了大鼠腹主动脉缩窄后左心室ＨＳＰ７０ｍＲＮＡ的水平。结果表明：大鼠腹主动脉缩窄后４ｈ,动脉血压已明显升高,并持续在高水平上;大鼠左心室重／体重比在第三天开始增加,然后持续升高,第４周时比对照组增加５９％;左心室ＨＳＰ７０ｍＲＮＡ在腹主动脉缩窄后４ｈ已明显升高,１ｄ,２ｄ,１ｗ均维持在高水平,１ｗ后逐渐消失。实验结果提示：大鼠心肌负荷增加早期,左心室ＨＳＰ７０ｍＲＮＡ表达明显增加。     

沙打旺组织培养中脱分化细胞的超微结构及细胞核的动态变化

对沙打旺的下胚轴、子叶、幼叶组织培养中脱分化细胞进行了超微结构观察,并着重讨论了细胞核的动态变化。脱分化细胞的细胞质中线粒体墙加,嵴明显;多聚核糖体增多;高尔基体增加;质体中积累淀粉。核仁与核内异染色质之间有一个动态过程。此过程暂称“核仁物质喷射“现象。在致有以下：１．核体出现,半嵌在增大的核仁上,核内异染色质沿核膜凝聚;２．异染色质移向核仁,并与核仁接触,核体消失,部分异质进入核仁;３．核仁物质     

Existence and Functions of Neurotens in Human Early Placental Villi

ＥｘｉｓｔｅｎｃｅａｎｄＦｕｎｃｔｉｏｎｓｏｆＮｅｕｒｏｔｅｎｓｉｎＨｕｍａｎＥａｒｌｙＰｌａｃｅｎｔａｌＶｉｌｌｉＺＨＡＮＧＣｈｏｎｇ－ｌｉ（张崇理）;ＣＨＥＮＧＬｉ－ｒｅｎ（程丽仁）,ＳＨＥＮＷｅｉ－ｂｉｎ（沈卫斌）;ＹＩＮＨｏｎｇ（殷红）;ＨＵ...     

大鼠TGF－α修饰16肽的溶液构象

已知大鼠Ｔ０Ｆ－α的抗原部位干Ｃ环,且大鼠的ＴＧＦ—α（３４－４３）和ＴＧＦ－α（４４－５０）均有较强的活性以转化正常细胞。为了揭示其结构与功能关系,合成了大鼠ＴＧＦ－α修饰１６肽。在前文用二维核磁共振技术归属质子谱并验证其一级结构的基础上,本文测定了不同混合时间的二维ＮＯＥＳＹ谱。根据所得的数据因原始斜率法求出距离约束。通过约束分子动力学计算定出该分子的溶液构象。其中还用ＤＱＦ－ＣＯＳＹ的数据求出二面角作为决定溶液构象的参考。     

cⅠ857基因的体外定位同义突变

 ｃⅠ８５７基因的体外定位同义突变陈南春,高辉,陈苏民,杨萍,刘新平（西安第四军医大学分子生物学研究所,西安７１００３２）外源基因要在大肠杆菌中获得高表达,需要合适的ＳＤ序列和可调控的强启动子［１］。ＰＬ启动子在原核启动子中属强启动子,它受ｃⅠ基因产物...