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71.
72.
Paul D. Roepe LiYong Wei Mary M. Hoffman Friederike Fritz 《Journal of bioenergetics and biomembranes》1996,28(6):541-555
Overexpression of the MDR protein, or p-glycoprotein (p-GP), in cells leads to decreased initial rates of accumulation and altered intracellular retention of chemotherapeutic drugs and a variety of other compounds. Thus, increased expression of the protein is related to increased drug resistance. Since several homologues of the MDR protein (CRP, ltpGPA, PDR5, sapABCDF) are also involved in conferring drug resistance phenomena in microorganisms, elucidating the function of the MDR protein at a molecular level will have important general applications. Although MDR protein function has been studied for nearly 20 years, interpretation of most data is complicated by the drug-selection conditions used to create model MDR cell lines. Precisely what level of resistance to particular drugs is conferred by a given amount of MDR protein, as well as a variety of other critical issues, are not yet resolved. Data from a number of laboratories has been gathered in support of at least four different models for the MDR protein. One model is that the protein uses the energy released from ATP hydrolysis to directly translocate drugs out of cells in some fashion. Another is that MDR protein overexpression perturbs electrical membrane potential () and/or intracellular pH (pHi) and therebyindirectly alters translocation and intracellular retention of hydrophobic drugs that are cationic, weakly basic, and/or that react with intracellular targets in a pHi, or -dependent manner. A third model proposes that the protein alternates between drug pump and Cl– channel (or channel regulator) conformations, implying that both direct and indirect mechanisms of altered drug translocation may be catalyzed by MDR protein. A fourth is that the protein acts as an ATP channel. Our recent work has tested predictions of these models via kinetic analysis of drug transport and single-cell photometry analysis of pHi, , and volume regulation in novel MDR and CFTR transfectants that have not been exposed to chemotherapeutic drugs prior to analysis. This paper reviews these data and previous work from other laboratories, as well as relevant transport physiology concepts, and summarizes how they either support or contradict the different models for MDR protein function. 相似文献
73.
利用人粒细胞集落刺激因子(hG-CSF)cDNA3′端非翻译区(3′-UTR)中存在的DraⅠ酶切位点,通过部分酶切与完全酶切,删除3′-UTR不同长度,构建了四种hG-CSFcDNA瞬时重组表达质粒。转染COS-7细胞后,生物活性测定结果提示,hG-CSFcDNA3′-UTR对其表达起负调控作用,其关键性序列位于紧接终止密码子TGA下游的65bp范围内,3′-UTR对hG-CSFcDNA表达的影响与转录水平的差别有一定关系。 相似文献
74.
We describe a new immunoassay which combines an immunosorbent assay, Immunogold silver staining and epipolarization microscopy. Our new assay procedure features multiple samples on a single microscope slide, and high sensitivity of epipolarization microscope for detection of silver-enhanced colloidal gold as a final immunoassay product. We call the new immunoassay “on slide immunogold assay” (OSIGA). This new method uses biotinylated antibody and streptavidin-gold reaction with silver enhancement technique. With OSIGA it is possible to investigate 30 samples on a single microscopic slide. Our preliminary studies used 10-20 μ1 samples and detected nanogram quantities of a standardized protein solution. Unlike enzyme linked immunosorbent assay (ELISA), which has a limited time for reading the final color products, the OSIGA specimens can be dried or resin mounted for longer storage and future reference. 相似文献
75.
76.
比较研究几种兼性和专一性CAM植物材料的PEPC同工酶表明:经自然干旱诱导,兼性CAM植物露花(Mesembryanthemumcordifolium)、长药景天(Sudumspectabile)有新的PEPC同工酶的出现,诱导前后各同工酶的天然分子量变化不大;而土三七(Sedumaizoon)则没有新的PEPC同工酶出现,但诱导后其同工酶的天然分子量有所增大。以上几种兼性CAM植物的PEPC同工酶酶谱无明显昼夜变化。专一性CAM植物的PEPC酶谱和天然分子量均较一致,亦无昼夜差异。 相似文献
77.
V. N. Kazakov P. Ya. Kravtsov I. E. Kuznetsov A. V. Tereshchenko 《Neurophysiology》1993,25(4):236-245
Responses of neurons of the lateral (LPO) and medial (MPO) subdivisions of the preoptic region (RPO) and of the supraoptic nucleus (SO) of the hypothalamus at infusions of up to 400 µl of a hypotonic (0.2%) or a hypertonic (0.3%) NaCl solution or an isotonic (5.5%) glucose solution into the homolateral internal carotid artery were studied in acute experiments on ketamine-anesthetized cats. Changes in the firing frequency were exhibited by 69% of the examinedRPO andSO neurons, the response being of four different types: a monophasic increase (1st-type) or a monophasic decrease (2nd-type) in the activity; biphasic responses where an initial frequency rise was followed by inhibition (3rd-type) and vice versa (4th-type). Of all the neuronal responses to all stimulations, 50% (121/245) were of the 1st type; 11% (26/245), of the 2nd type; 5% (14/245), of the 3rd type; and 3% (7/245), of the 4th type. No neurons with 1st-type responses to infusion of the hypotonic NaCl solution were found in the medial sections of theMPO, and of the hypertonic solution, in the lateral sections of theLPO. Neurons with 2nd-type responses to infusion of hypo- and hypertonic NaCl solutions were almost solely detected in theSO, whereas units with such responses to glucose infusions were observed only in theRPO, being fully absent in theSO, where this stimulation gave rise only to 1st-type responses. Neurons with 3rd- and 4th-type responses to hypo- and hyperosmotic stimulations were detected predominantly in theMPO andSO, and to glucose stimulation, in theMPO andLPO.Neirofiziologiya/Neurophysiology, Vol. 25, No. 4, pp. 281–291, July–August, 1993. 相似文献
78.
腹腔注射链脲佐霉素(65mg/kg)诱发Wistar大鼠糖尿病。糖尿病发病4周后,向饲料中加尼群地平(30mg·kg-1·d-1)。结果表明,糖尿病4周时大鼠心室舒张功能首先受损,8周后心室舒张和收缩功能均明显受累。尼群地平处理对糖尿病大鼠的心肌收缩性有一定的改善作用。提示尼群地平对大鼠糖尿病性心肌病有一定有益作用。 相似文献
79.
Through simple model analysis, the mass action kinetic model for lipolytic enzymes in biphasic aqueous-organic systems can be simplified using the quasi-steady state assumption (or the quasi-equilibrium state assumption) for the adsorbed enzyme E* or the enzyme-substrate complex E*S. Some parameter combinations leading to the above assumptions are derived confirmed by full numerical integration of the whole enzymatic process. The results may be classified into three categories: (1) the quasi-equilibrium state assumption for E*, (2) the quasi-steady state assumption for E*, and (3) the quasi-steady state assumption for E*S. Further simplification for both E* and E*S is also discussed. (c) 1993 Wiley & Sons, Inc. 相似文献
80.