Inhibitory Monoclonal Antibodies and Their Recombinant Derivatives Targeting Surface-Exposed Carbonic Anhydrase XII on Cancer Cells

Monoclonal and recombinant antibodies are widely used for the diagnostics and therapy of cancer. They are generated to interact with cell surface proteins which are usually involved in the development and progression of cancer. Carbonic anhydrase XII (CA XII) contributes to the survival of tumors under hypoxic conditions thus is considered a candidate target for antibody-based therapy. In this study, we have generated a novel collection of monoclonal antibodies (MAbs) against the recombinant extracellular domain of CA XII produced in HEK-293 cells. Eighteen out of 24 MAbs were reactive with cellular CA XII on the surface of live kidney and lung cancer cells as determined by flow cytometry. One MAb 14D6 also inhibited the enzymatic activity of recombinant CA XII as measured by the stopped-flow assay. MAb 14D6 showed the migrastatic effect on human lung carcinoma A549 and renal carcinoma A498 cell lines in a ‘wound healing’ assay. It did not reduce the growth of multicellular lung and renal cancer spheroids but reduced the cell viability by the ATP Bioluminescence assay. Epitope mapping revealed the surface-exposed amino acid sequence (35-FGPDGENS-42) close to the catalytic center of CA XII recognized by the MAb 14D6. The variable regions of the heavy and light chains of MAb 14D6 were sequenced and their complementarity-determining regions were defined. The obtained variable sequences were used to generate recombinant antibodies in two formats: single-chain fragment variable (scFv) expressed in E. coli and scFv fused to human IgG1 Fc fragment (scFv-Fc) expressed in Chinese Hamster Ovary (CHO) cells. Both recombinant antibodies maintained the same specificity for CA XII as the parental MAb 14D6. The novel antibodies may represent promising tools for CA XII-related cancer research and immunotherapy.     

稠油微生物开采技术现状及进展

综述了用微生物方法开采稠油的技术现状与进展，论题如下。①概述。②基本方法：异源微生物采油，包括微生物吞吐和微生物驱；本源微生物采油压大港孔店油田的实例。③主要机理，包括产表面活性剂，降解稠油中重质组分及其他。④技术研究，包括机理性、可行性及经济效益研究，列举了国内外6个实例。⑤现场应用，包括国外1个、国内6个实例。⑥该技术的优势及问题。参22。     

Random errors caused by temperature in magnitude of principal strains evaluated ith 3-element strain gauge rosettes

The effects of the uncertainties associated ith the apparent strain and gauge factor data, given by the manufacturer, in the evaluation of principal strains are examined. Principal strains are obtained by a reduction of the strains measured ith electrical resistance strain gauge rectangular or delta rosettes. The theoretical analysis points out the relevance of the effects caused.     

Thermomechanical Models of Irreversible Finite Deformation of Anisotropic Solids

The proposed models for irreversible finite deformation of initially anisotropic solids are based on the hypothesis of the parameters of state, for which the authors have chosen a universal parameter (temperature) and a strain component, which is analogous to volume strains in an isotropic body and represents thermal strains of an unconstrained (stress-free) anisotropic solid. The application of this hypothesis allows the strain tensor to be resolved into a reversible and irreversible components. For irreversible processes of finite deformation of hardenable materials, the authors have proposed a deformation-type model, which makes it possible to describe a change in the type of anisotropy of initially anisotropic materials in the course of deformation. For the flow-theory-type model, the limiting state function, as given in the space of irreversible strains, allows for the experimentally observed plastic flow of anisotropic crystals under the action of hydrostatic pressure only.     

农业重组微生物生物安全研究进展

综述了近年来农业重组微生物的研究现状和进展 ,详细介绍农业重组微生物环境释放的监控方法以及对环境的冲击作用 ,对一些有关生物安全研究的热点问题进行了探讨     

Application of artificial neural networks to the analysis of two‐dimensional fluorescence spectra in recombinant E coli fermentation processes

A two‐dimensional (2D) spectrofluorometer was used to monitor various fermentation processes with recombinant E coli for the production of 5‐aminolevulinic acid (ALA). The whole fluorescence spectral data obtained during a process were analyzed using artificial neural networks, ie self‐organizing map (SOM) and feedforward backpropagation neural network (BPNN). The SOM‐based classification of the whole spectral data has made it possible to qualitatively associate some process parameters with the normalized weights and variances, and to select some useful combinations of excitation and emission wavelengths. Based on the classified fluorescence spectra a supervised BPNN algorithm was used to predict some of the process parameters. It was also shown that the BPNN models could elucidate some sections of the process's performance, eg forecasting the process's performance. Copyright © 2005 Society of Chemical Industry     

Effect of temperature on recombinant protein production using the Bm5/Bm5.NPV expression system

A series of experiments have been conducted using a recombinant baculovirus/insect cell expression system (Bm5/Bm5.NPV.CAT) to establish the optimum temperature for both cell growth and virus infection. Bm5 cell growth was found to be limited at temperatures below 22°C and ceased completely at temperatures above 34°C. In the range between 24 and 28°C, final cell densities always reached 96% of the highest achievable viable cell density. The shortest population doubling time was obtained at 28°C. Overall, a consistent increase in metabolism with increasing temperatures was observed. During the infection/viral replication phase, an increase in the temperature from 25 to 31°C resulted in a faster decrease in viable cell density and an earlier production of chloramphenicol acetyltransferase (CAT). Furthermore, protein yield at temperatures above 28°C was significantly reduced. Overall, the best temperature for the infection phase for the Bm5/Bm5.NPV expression system was found to be 25°C when the cells are cultured in serum free media.     

微生物法处理含硫工业废气

指出了含硫工业废气传统处理方法的缺陷 ,介绍了微生物处理方法的优势所在 ;对微生物菌种的共同特征以及无机硫和有机硫脱除时菌种的选择依据进行了讨论 ,并对微生物法脱硫的机理进行了分析 ;评述了国内外生物滤池法、生物滴滤池法和生物吸收法等生物膜法脱硫的工艺及其工业化应用。指出了微生物法脱硫技术有待深入研究的问题、发展趋势和应用前景。     

Ethanol production from raw starch by a recombinant yeast having saccharification and fermentation activities

In order to develop a method for converting raw starch into ethanol efficiently, direct fermentation of ozonized raw starch using a recombinant yeast was investigated. Ozonolysis was carried out as a pretreatment to convert raw starch into ethanol rapidly and efficiently, and then the effect of the ozone degradation conditions on the degree of polymerization and the amount of amylose in a raw starch was determined. Since the degree of polymerization was low and the amount of amylose was high, raw starch treated with an ozone concentration of 40 gm^?3 and an ozonation time of 30 min was the material chosen for alcohol fermentation. Though the recombinant yeast could not convert the untreated raw starch, it converted the soluble starch and the ozonized raw starch at a comparatively high yield into ethanol. About 56% of the ozonized raw starch decomposed, and the ethanol concentration obtained from the ozonized raw starch was markedly greater than that obtained from untreated raw starch. The dynamic behavior of cell growth, substrate degradation, and ethanol production was examined in a continuous culture under various dilution rates, and the optimal dilution rate, ie 0.15 h^?1, was determined for maximizing the ethanol productivity (amount of ethanol produced per unit time). © 2002 Society of Chemical Industry