The effects of induction conditions on production of a soluble antitumor sFv in Escherichia coli

CC49 is a second generation monoclonal antibody(mAb) with highaffinity to a pancarcinoma antigen, TAG-72. A single-chain Fv(sFv)ofCC49 may have a role in managing human carcinomas. Most reportedsFvs have been expressed as insoluble products that must besolubilized and renatured. Soluble sFv expression is advantageousas activity can be assayed directly from the periplasmic fraction.Also, gene-level immunoconjugates may not be amenable to refoldingprotocols. Using a vector that carries the tac promoter andomp A signal, we have examined the effects of four variableson the expression and accumulation of soluble CC49 sFv: (i)linker sequence joining VL and VH, (ii) isopropylthio-ß-D-galactosideconcentration for induction, (iii) temprature, and (iv) theaddition of nonmetabolizable sugars to the medium. We have beenable to demonstrate, using rapidly prepared periplasmic extracts,that the yield of soluble sFv improves by the addition of 0.4Msucrose to the medium and by inducing expression with a verylow concentration of IPTG (0.02–0.03 mM). Under theseinduction conditions periplasmic extracts demonstrate increasedexpression of the sFv, as shown by the larger amount of a 27kDa band on SDS-polyacrylamide gel, and an increased abilityto inhibit binding of the mAb CC49 to immobilized tumor extracts.     

Automating the identification and analysis of protein {beta}-barrels

ßBarrels are widespread and well-studied featuresof a great many protein structures. In this paper an unsuper-visedmethod for the detection of P-barrels is developed based ontechniques from graph theory. The hydrogen bonded connectivityof ß-sheets is derived using standard pattern recognitiontechniques and expressed as a graph. Barrels correspond to topologicalrings in these connectivity graphs and can thus be identifiedusing ring perception algorithms. Following from this, the characteristictopological structure of a barrel can be expressed using a novelform of reduced nomenclature that counts sequence separationsbetween successive members of the ring set These techniquesare tested by applying them to the detection of barrels in anon-redundant subset of the Brookhaven database. Results indicatethat topological rings do seem to correspond uniquely to ß-barrelsand that the technique, as implemented, finds the majority ofbarrels present in the dataset.     

吲哚美辛对荷瘤小鼠抗肿瘤作用机制的实验研究

用微观放射自显影技术及免疫组化法探讨了非甾体类抗炎药吲哚美辛(IN)对Lewis肺癌的生长抑制作用机制。结果显示:与对照组相比,实验组IN的抑瘤率为55.05%;IN可显著下调Bcl-2蛋白的表达(t=6.154,P<0.001),对Bax蛋白的表达没有明显影响(t=2.101,P>0.05)。放射自显影结果显示,在肿瘤细胞的细胞膜、细胞浆及细胞核中均有黑色银颗粒分布,且在用药后12 h时肿瘤组织中的平均银颗粒数高于4、7、243、6 h时。以上结果表明,IN可抑制Lewis肺癌的生长,其机制与下调抗凋亡蛋白Bcl-2的表达及降低Bcl-2/Bax的比值有关;IN可进入到肿瘤细胞内发挥抗肿瘤作用。     

Nutritional response of mature rats to kidney bean (Phaseolus vulgaris) lectins

In a series of 10-day pair feeding experiments it was found that the nutritional value of diets containing beans was essentially the same for rats aged between 30 and 123 days. Thus net protein utilisation (NPU) values of 25–39 on diets containing Processor bean (35 g protein kg^?1) + egg albumin (65g protein kg^?1) were obtained. As food intakes were considerably reduced when rats were fed diets containing more than 35g protein kg^?1 of Processor bean, the measurement of protein utilisation became increasingly more difficult. The severe disruption of the brush borders of duodenal and jejunal enterocytes, originally observed when bean-containing diets were fed to young (30-day-old) rats was also found with rats up to the age of 120 days on similar diets. Similarly, the development of circulating anti-lectin antibodies in the animals showed no age dependence within the age limits investigated. It was also shown that oral immunisation did not protect the rats from the effects of toxicity and that the immune response was a result of continuous absorption of lectin throughout the feeding period. Thus the extent and the mechanism of toxicity of Phaseolus vulgaris bean lectins were found not to be dependent on the age or maturity of the animal.     

Paste Residence Time in a Spouted Bed Dryer. II: Effect of Spout Operational Conditions

Residence time distributions for an aqueous solution of 10% sodium chloride in a spouted bed dryer of inert particles were determined using the stimulus-response technique. Glass and polyethylene beads with diameters 2.6 and 3.4 mm were used as inert bodies in a cylindrical column of 14.0 cm diameter and 60° conical base. The effects of inert bodies load, air, and paste flow rates on the mean residence times and RTD were determined following 2³ factorial designs. The RTD could be correlated to the perfect mixing cell model with R² varying from 0.8684 to 0.9815. The mean residence times in CSBD varied from 10.8 to 13.9 and 10.7 to 13.3 min for glass and polyethylene beads, respectively. For both inert particles, mean residence times increased with bed height and decreased with paste feed rates. Also, terms of interaction among the factors were significant in some cases, showing a complex behavior of paste residence times. Equations obtained by response surface regression could predict mean residence times on glass and polyethylene beads with deviations lower than ±10%.     

高压锅炉补给水系统管道的腐蚀

讨论了锅炉补给水管道的腐蚀情况 ,指出溶解氧和二氧化碳是产生腐蚀的主要原因 ,其腐蚀速率为0 .4 2mm/a。确保真空除气系统正常运行和二级除盐水箱的密封效果是控制腐蚀的主要手段。在此情况下 ,该管线尚可运行 7年     

应用底饲回燃循环流化床技术改造东锅DG35/3.82-7型流化床锅炉

采用飞灰循环底饲回燃技术改造西南合成制药总厂热电分厂、天府矿务局磨心坡发电厂2台东方DG35/3.82-7型低倍率循环流化床锅炉。改造后运行结果表明,锅炉可长期、稳定运行,尾部飞灰可燃物含量大幅度降低,锅炉蒸发量提高20％左右,节煤效果明显。为燃用贫煤、无烟煤和煤矸石等劣质燃料且锅炉炉膛高度较低的电站技改,提供了一种切实可行的方法。     

环冷机卸矿槽料位自动控制技术的改进

针对烧结过程中环冷卸矿槽料位存在周期性变化的特点。提出了一种改进的控制思路和解决方案。该方案在太钢三烧450m^2烧结机上采用后，取得了较好的效果。     

Drying of Probiotics: Optimization of Formulation and Process to Enhance Storage Survival

This work describes how probiotic bacteria can be dried at low temperature in two steps, combining spray drying and vacuum drying, in order to enhance their survival during storage. A sufficient number of dried probiotics survived storage for more than 3 months at 30°C, if an appropriate combination of protein and carbohydrate was selected as carrier and storage conditions were maintained optimal. The use of soy protein and maltodextrin or skim milk and arabic gum resulted in the best survival rates of probiotics during storage. No evident difference was found between different spray dryer configurations, although a cocurrent flow was preferred.     

V. Yeast sequencing reports. Identification of a gene encoding a new Ypt/Rab-like monomeric G-protein in Saccharomyces cerevisiae

A Saccharomyces cerevisiae sequence cloned by serendipity was found to encode a protein that is a new member of the Ypt/Rab monomeric G-protein family. This sequence shows high homology to the yeast genes SEC4 and YPT1 and, like SEC4 and YPT1, is essential for viability. The sequence was localized to chromosome V based upon hybridization to pulse-field gel-separated yeast chromosomes. The sequence has been deposited in the GenBank data library under Accession Number L17070.