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101.
    
The cytotoxicity of a recombinant interleukin 6 (IL-6)-diphtheria toxin (DT) fusion protein towards human myeloma cell lines was investigated. DAB389-IL-6 inhibited protein synthesis and methylcellulose colony formation by U266 myeloma cells. In the clonogenic assay, the fusion protein approached the level of cytotoxicity achieved by native DT. The specificity of killing by DAB389-IL-6 was demonstrated by inhibition of cytotoxicity by a molar excess of free rhIL-6. The effect of DAB389-IL-6 on colony formation by six OCI-My cell lines was assessed. Similar to U266 cells, colony growth by the OCI-My 5 and -My 2 cell lines was inhibited in a simple dose dependent manner. However, a biphasic effect was observed for the IL-6 dependent OCI-My 4 cells; DAB389-IL-6 stimulated colony formation at low (< or = 10(-11) M) concentrations, yet was inhibitory at higher doses. Three other cell lines whose growth was not altered by IL-6 were relatively unaffected by DAB389-IL-6, despite their sensitivity to native DT. Flow cytometric analysis for IL-6 receptor expression using phycoerythrin-conjugated IL-6 demonstrated specific binding sites on both DAB389-IL-6 sensitive and certain insensitive cell lines, suggesting that other factors in addition to the expression of IL-6 receptors are involved in killing by the fusion toxin. Despite evidence for a role of IL-6 in myeloid cell development, normal bone marrow was insensitive to the IL-6 fusion toxin. In cultures containing both normal bone marrow and U266 cells DAB389-IL-6 effectively inhibited the growth of U266 myeloma colonies but had little effect on normal bone marrow erythroid, granulocyte and mixed erythroid/granulocyte colony growth. From these experiments we conclude that DAB389-IL-6 is specifically cytotoxic towards a subset of IL-6-responsive human myeloma cell lines and may be useful, in some cases, in the selective elimination of tumour cells from mixed populations of normal and malignant cells.  相似文献   
102.
    
The hormone-binding domain of the glucocorticoid receptor must be bound to heat shock protein (hsp) 90 for it to have a high-affinity steroid-binding conformation. Cell-free assembly of a glucocorticoid receptor-hsp90 heterocomplex is brought about in reticulocyte lysate by a preformed protein-folding complex containing hsp90, hsp70, and other proteins [Hutchison, K.A., Dittmar, K. D., & Pratt, W.B. (1994) J. Biol. Chem. 269, 27894-27899]. In this \"foldosome\" system, hsp70 is required for assembly of the receptor-hsp90 complex and concomitant activation of steroid-binding activity [Hutchison, K.A., Dittmar, K.D., Czar, M.J., & Pratt, W.B. (1994) J. Biol. Chem. 269, 22157-22161]. All previous experiments involving cell-free assembly of both receptor-hsp90 and protein kinase-hsp90 heterocomplexes have been carried out with the protein-folding system in rabbit reticulocyte lysate. In this work, we show that concentrated lysates of receptor-free mouse (L cells) and insect (Sf9) cells and also a plant (wheat germ) lysate fold the immunopurified glucocorticoid receptor into a functional (i.e., steroid binding) heterocomplex with hsp90. Receptor heterocomplex formation in animal lysates and in the plant lysate are not identical in that the dynamics of complex assembly are different, but both systems produce a functional complex that binds steroid. Also, in contrast to animal and insect complexes, receptor-plant hsp90 complexes are not stabilized by molybdate. When added to the other lysate, purified plant and animal hsp90s show partial complementarity, in that a receptor-hsp90 complex is formed but the receptor is not converted to the steroid-binding conformation. When added to rabbit reticulocyte lysate that has been depleted of endogenous hsp70, purified wheat germ and mouse hsp70's are equally active in promoting both assembly of receptor-hsp90 heterocomplexes and conversion of receptor to the steroid-binding conformation. Thus, hsp70 from the plant kingdom has conserved the ability to interact functionally with chaperone proteins of the animal kingdom to cooperate in protein folding as evidenced by formation of a functional receptor-hsp90 heterocomplex.  相似文献   
103.
    
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104.
    
The conjugation of 1-p-chlorophenyl-4,4-dimethyl-5-diethylamino-1-penten-3- one hydrobromide (CDDP), a Mannich base of an alpha,beta-unsaturated ketone, to glutathione is catalyzed selectively by alpha-class glutathione S-transferase. The reaction of CDDP with glutathione can be monitored continuously by following the conjugation-dependent decrease in absorbance of CDDP at 307 nm. The Km and Vmax for CDDP with alpha-class glutathione S-transferase from horse liver were determined to be 226 microM and 14.6 mumol/(min.mg), respectively. CDDP is the first example of an alpha-class glutathione S-transferase selective substrate that monitors the glutathione conjugation activity, rather than the glutathione peroxidase activity of the enzyme.  相似文献   
105.
THE DESIGN OF EXPERIMENTS FOR SHELF LIFE STUDY   总被引:13,自引:0,他引:13  
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106.
107.
The finite element method was used to determine the stress distributions in adhesive joints prepared using alumina adher-ends and silicate glass adhesives with varying thicknesses and thermal expansion coefficients. These analyses, together with the results of literature analyses for joints subjected to externally applied loads, aided in interpreting the experimental observations which are described in Part II of the paper.  相似文献   
108.
The subsolidus compatibility relations in the system SrO-B2O3- SiO2 were determined by solid-state reaction techniques and X-ray powder diffraction methods. The system was found to contain 11 subsolidus compatibility relations, one stable ternary compound (Sr3B2SiO8), and one metastable ternary compound with a probable composition SrB2Si2O8.  相似文献   
109.
Phytic Acid Determination in Soybeans   总被引:4,自引:0,他引:4  
Several established methods of phytic acid determination in soybeans were evaluated. Iron analysis methods, which rely on a 4:6 molar ratio of Fe:P, were eliminated because this ratio was not dependable. Three assay methods relying on phosphorus analysis were then compared. The anion-exchange method was considered most accurate but not convenient for routine analysis. Analysis of the ferric phytate precipitate and a new method, analysis of the supernatant before and after ferric chloride precipitation, were judged against the anion-exchange method under different extraction conditions. Based upon good agreement with anion-exchange column results and acceptable reproducibility, the best methods were (1) precipitate analysis of phosphorus after extraction with 3% TCA + 10% sodium sulfate, or (2) the supernatant difference method after extraction with 1.2% HCl.  相似文献   
110.
An analysis of dilute, turbulent particle/liquid two-phase flow is presented. The three-dimensional conservation equations that govern turbulent motion in solid/fluid flows are derived using ensemble averaging, and the unknown terms in these equations are constituted to achieve closure. These closure terms include the shear stress due to interparticle collisions, the corresponding terms in the Reynolds stress equation, the force and dissipation due to particle-wall collisions, and the interfacial work due to particle/turbulent eddy interaction. The resultant two-fluid model was then evaluated using a computational fluid dynamic (CFD) solver and the predictions were compared with experimental data. Good agreement was observed for a variety of flow conditions.  相似文献   
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