Direct antigen capture by soluble scFv antibodies

For ease of detection, soluble forms of phage-displayed scFv antibodies are usually expressed with a tag, e.g., c-myc or His (Histidine). The binding is then assayed by a monoclonal antibody to the tag. In this article, we describe the use of biotinylated antigen for detecting soluble scFv antibodies without utilizing the peptide tag detection system. The scFv antibodies were against the oncoplacental antigen heat-stable alkaline phosphatase (HSAP). The method essentially consisted of either reverse Western or antigen capture enzyme-linked immunosorbent assay (ELISA). In the reverse Western, periplasmic extract was electrophoresed, and binding to biotinylated antigen was detected by the detection system based on streptavidin-horseradish peroxidase. The antigen capture ELISA utilized the binding of periplasmic extract to a polystyrene plate. We have also demonstrated the use of antigen capture ELISA for studying specificity and affinity of the selected clones. Although these techniques have been developed for antibodies to HSAP, they have general utility for phage expression systems without a peptide tag.     

Differential staining of Western blots of human secreted glycoproteins from serum, milk, saliva, and seminal fluid using lectins displaying diverse sugar specificities

Human milk, serum, saliva, and seminal fluid glycoproteins (gps) nourish and protect newborn and adult tissues. Their saccharides, which resemble cell membrane components, may block pathogen adhesion and infection. In the present study, they were examined by a battery of lectins from plants, animals, and bacteria, using hemagglutination inhibition and Western blot analyses. The lectins included galactophilic ones from Aplysia gonad, Erythrina corallodendron, Maclura pomifera (MPL), peanut, and Pseudomonas aeruginosa (PA-IL); fucose-binding lectins from Pseudomonas aeruginosa (PA-IIL), Ralstonia solanacearum (RSL), and Ulex europaeus (UEA-I), and mannose/glucose-binding Con A. The results demonstrated the chosen lectin efficiency for differential analysis of human secreted gps as compared to CBB staining. They unveiled the diversity of these body fluid gp glycans (those of the milk and seminal fluid being highest): the milk gps interacted most strongly with PA-IIL, followed by RSL; the saliva gps with RSL, followed by PA-IIL and MPL; the serum gps with Con A and MPL, followed by PA-IIL and RSL, and the seminal plasma gps with RSL and MPL, followed by UEA-I and PA-IIL. The potential usage of these lectins as probes for scientific, industrial, and medical purposes, and for quality control of the desired gps is clearly indicated.     

Application of exploratory data analysis for the characterization of tubular wells of the North of Brazil

Roraima is located in the extreme North of Brazil, in the Western Amazon Region. The groundwater has an important role in the public supply of potable water on the Roraima State. The objective of this work is the application of PCA (Principal Component Analysis) and HCA (Hierarchical Cluster Analysis) for the characterisation of tubular wells of the Roraima State, as regards physical–chemical composition of water. The parameters analyzed in laboratory were the following ones: pH, electric conductivity, Total Hardness, Total Alkalinity, Bicarbonate, Calcium, Potassium, Magnesium, Nitrate, Chloride, Total Iron. The physical–chemical analyses, were carried out during the months of September and October 2000 (winter, period of rains). The laboratory analysis was performed using classical methods (titrimetry and uv–visible spectroscopy), the samples were processed in agreement with the Standard Methods for Examination of Water and Wastewater.The physical–chemical analyses, were accomplished in September and October 2000. In physical–chemical terms, the groundwater samples analyzed do not indicate any restriction to human consumption and the two chemometric methods of exploratory data analysis: PCA and HCA are efficient for the discrimination of bicarbonated (tubular wells locateds in the South of the Roraima State) and not bicarbonated (tubular wells locateds in the North of Roraima) groundwater.     

Comprehensive study on saline-alkali soil amelioration with sediment of irrigation area in northeast China

To improve the main shortcomings of insufficient nutrients, high salinity and low productivity in saline-alkali soils, this study collected saline-alkali soil samples from Da'an City of Baicheng and sediment samples from the adjacent Qianguo irrigation district, measured their pH values and major element contents, designed and verified the improvement scheme of saline-alkali soil based on geochemical engineering principles. Results indicated that the pH value and various element contents of saline-alkali soil could be adjusted by adding sediment to make it close to reference soil. Both theoretical calculation and experimental determination (soil pH, mineral composition, functional group composition and crop growth) revealed that the optimal mixing ratio of saline-alkali soil and sediment was 2:1. The saline-alkali soil amelioration with sediment of adjacent irrigation areas was effectively, low-cost and environment-friendly. The geochemical engineering principle was applicable to the amelioration of saline-alkali soil, but the mixing ratio of saline-alkali soil and sediment in different areas should vary with the local conditions. Meanwhile, theoretical calculation could basically replace the experimental determination to simplify the actual engineering application process. This saline-alkaline soil amelioration method makes full use of the surrounding natural waste and has the advantage of adapting measures to local conditions.     

Effects of Angiotensin II on Erythropoietin Production in the Kidney and Liver

The kidney is a main site of erythropoietin production in the body. We developed a new method for the detection of Epo protein by deglycosylation-coupled Western blotting. Detection of deglycosylated Epo enables the examination of small changes in Epo production. Using this method, we investigated the effects of angiotensin II (ATII) on Epo production in the kidney. ATII stimulated the plasma Epo concentration; Epo, HIF2α, and PHD2 mRNA expression in nephron segments in the renal cortex and outer medulla; and Epo protein expression in the renal cortex. In situ hybridization and immunohistochemistry revealed that ATII stimulates Epo mRNA and protein expression not only in proximal tubules but also in collecting ducts, especially in intercalated cells. These data support the regulation of Epo production in the kidney by the renin–angiotensin–aldosterone system (RAS).     

西太平洋海山玄武岩等离子体质谱分析及其地质意义

海山玄武岩记录着海山演化以及相关深部过程的重要信息。对西太平洋戈沃罗夫平顶海山、牛郎平顶海山、阿诺德平顶海山、斯科尔尼亚科瓦平顶海山、戈尔金平顶海山、麦克唐奈平顶海山、令仪海山、徐福平顶海山玄武岩进行岩石学和等离子体质谱测试分析,结果表明:研究区玄武岩多呈杏仁状、气孔-杏仁状、块状构造,斑状结构。岩石主要由斑晶、基质组成。斑晶的矿物成份主要为斜长石、橄榄石和单斜辉石。基质的矿物成份主要为斜长石、橄榄石、单斜辉石和玻璃质等。不同海山玄武岩的稀土配分模式以及稀土参数均反映了洋岛玄武岩的典型特征,表明了研究区海山是一组属于多热点成因的典型板内火山型海山,可能受过了多期次的区内断裂岩浆活动的影响。     

遗传算法结合偏最小二乘法无损评价西洋梨糖度

基于遗传算法的波段选择法在组合优化问题上具有很大的搜索优势,适应性很广。文章将该方法应用于西洋梨糖度近红外光谱分析中,探讨数据优化筛选的可行性。光谱经多元散射校正或标准归一化处理后进行波段选择,选择结果与样品中被测成分有关, 4个品种洋梨的最佳个体染色体编码有一定共性。分别建立了四种洋梨的GA-PLS模型和全谱模型,早红考密斯、五九香、凯斯凯德和康佛伦斯的GA-PLS建模数据点分别从1 557减少到了434,496,310和496。GA-PLS/Fr-PLS模型的预测标准偏差分别为0.428/0.518,0.696/0.694,0.425/0.421和0.567/0.633, 其中早红考密斯和康佛伦斯GA-PLS模型的预测精度明显优于全谱模型,而五九香和凯斯凯德的GA-PLS模型与全谱模型相近。结果表明,遗传算法用于PLS建立西洋梨糖度校正模型前的数据优化筛选是可行的, 有效提高测量精度, 减少建模变量。     

Expression of Luteinizing Hormone-Releasing Hormone (LHRH) and Type-I LHRH Receptor in Transitional Cell Carcinoma Type of Human Bladder Cancer

Bladder cancer (BC) is the tenth most frequently detected cancer in both sexes. Type-I luteinizing hormone-releasing hormone (LHRH) receptor (LHRH-R-I) is expressed not only in the pituitary, but also in several types of cancer disease. There are few data about LHRH-R-I expression in human BC. This study aimed to investigate the expression of LHRH and LHRH-R-I in the transitional cell carcinoma (TCC) type of human BC. RNA was extracted from 24 human bladder tumor specimens and three BC cell lines. RT-PCR was performed to detect mRNA for LHRH and LHRH-R-I. The protein of LHRH-R-I was further studied by immunohistochemistry (IHC), ligand competition assay, and Western Blot. PCR products of LHRH were found in 19 of 24 (79%) specimens and mRNA of LHRH-R-I was detected in 20 of 24 specimens (83%). Positive immunostaining for LHRH-R-I with different expression intensity was found in all samples examined, showing negative correlation with TCC grade. Radioligand binding studies also showed the presence of specific LHRH-R-I and high affinity binding of LHRH analogs. The high incidence of LHRH-R in BC suggests that it could serve as a molecular target for therapy of human BC with cytotoxic LHRH analogs or modern powerful antagonistic analogs of LHRH.     

西部地区交通建设中的泥石流灾害与防治对策

我国西部地区铁路、公路交通干线共有泥石流灾害 4 70 0余处, 占全国道路泥石流灾害的 70 %以上。泥石流分布广泛, 成灾方式多样, 灾害严重, 制约了西部地区交通建设的发展。本文提出了做好路线方案比选, 防患于未然, 正确评判泥石流沟和发育现状, 工程设计要适合泥石流特点, 以及提高防灾减灾勘察设计水平, 开展泥石流防治试验示范工程研究等六条防治对策。