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111.
Effect of Hypoxia on Na+-K+-Cl− Cotransport in Cultured Brain Capillary Endothelial Cells of the Rat
Abstract: The effect of hypoxia on Na+,K+-ATPase and Na+-K+-Cl? cotransport activity in cultured rat brain capillary endothelial cells (RBECs) was investigated by measuring 86Rb+ uptake as a tracer for K+. RBECs expressed both Na+,K+-ATPase and Na+-K+-Cl? cotransport activity (4.6 and 5.5 nmol/mg of protein/min, respectively). Hypoxia (24 h) decreased cellular ATP content by 43.5% and reduced Na+,K+-ATPase activity by 38.9%, whereas it significantly increased Na+-K+-Cl? cotransport activity by 49.1% in RBECs. To clarify further the mechanism responsible for these observations, the effect of oligomycin-induced ATP depletion on these ion transport systems was examined. Exposure of RBECs to oligomycin led to a time-dependent decrease of cellular ATP content (by ~65%) along with a complete inhibition of Na+,K+-ATPase and a coordinated increase of Na+-K+-Cl? cotransport activity (up to 100% above control values). Oligomycin augmentation of Na+-K+-Cl? cotransport activity was not observed in the presence of 2-deoxy-d -glucose (a competitive inhibitor of glucose transport and glycolysis) or in the absence of glucose. These results strongly suggest that under hypoxic conditions when Na+,K+-ATPase activity is reduced, RBECs have the ability to increase K+ uptake through Na+-K+-Cl? cotransport. 相似文献
112.
A combined physical and genetic map of theCorynebacterium glutamicum ATCC 13032 chromosome was constructed using pulsed-field gel electrophoresis (PFGE) and hybridizations with cloned gene probes. Total genomic DNA was digested with the meganucleasesSwaI (5-ATTTAAAT-3),PacI (5-TTAATTAA-3), andPmeI (5-GTTTAAAC-3) yielding 26, 27, and 23 fragments, respectively. The chromosomal restriction fragments were then separated by PFGE. By summing up the lengths of the fragments generated with each of the three enzymes, a genome size of 3082 +/- 20 kb was determined. To identify adjacentSwaI fragments, a genomic cosmid library ofC. glutamicum was screened for chromosomal inserts containingSwaI sites. Southern blots of the PFGE gels were hybridized with these linking clones to connect theSwaI fragments in their natural order. By this method, about 90% of the genome could be ordered into three contigs. Two of the remaining gaps were closed by cross-hybridization of blottedSwaI digests using as probesPacI andPmeI fragments isolated from PFGE gels. The last gap in the chromosomal map was closed by hybridization experiments using partialSwaI digestions, thereby proving the circularity of the chromosome. By hybridization of gene probes toSwaI fragments separated by PFGE about 30 genes, including rRNA operons, IS element and transposon insertions were localized on the physical map. 相似文献
113.
Radioadaptive response: Efficient repair of radiation-induced DNA damage in adapted cells 总被引:13,自引:0,他引:13
To verify the hypothesis that the induction of a novel, efficient repair mechanism for chromosomal DNA breaks may be involved in the radioadaptive response, the repair kinetics of DNA damage has been studied in cultured Chinese hamster V79 cells with single-cell gel electrophoresis. The cells were adapted by priming exposure with 5 cGy of γ-rays and 4-h incubation at 37°C. There were no indication of any difference in the initial yields of DNA double-strand breaks induced by challenging doses from non-adapted cells and from adapted cells. The rejoining of DNA double-strand breaks was monitored over 120 min after the adapted cells were challenged with 5 or 1.5 Gy, doses at the same level to those used in the cytogenetical adaptive response. The rate of DNA damage repair in adapted cells was higher than that in non-adapted cells, and the residual damage was less in adapted cells than in non-adapted cells. These results indicate that the radioadaptive response may result from the induction of a novel, efficient DNA repair mechanism which leads to less residual damage, but not from the induction of protective functions that reduce the initial DNA damage. 相似文献
114.
Hisao Kurazono Shinji Yamasaki Orn-anong Ratchtrachenchai G. Balakrish Nair Yoshifumi Takeda 《Microbiology and immunology》1996,40(4):303-305
Vibrio cholerae O139 isolated from different countries, as well as from different locations within a country, were examined using macrorestriction DNA analysis to determine the clonality of the O139 strains. NotI digests of genomic DNA of representative strains from Nepal, India, Bangladesh, China, Thailand, and Malaysia revealed very similar but not identical patterns. Examinations of the banding patterns generated by pulsed-field gel electrophoresis of strains isolated within countries revealed complete homogeneity. These results further reiterate the spread of an identical clone of V. cholerae O139 although it appears that genetic polymorphism among the O139 strains is becoming apparent. 相似文献
115.
Kazushige Morimoto Eisuke Tsuda Ahmed Abdu Said Eriko Uchida Satoshi Hatakeyama Masatsugu Ueda Takao Hayakawa 《Glycoconjugate journal》1996,13(6):1013-1020
Ten erythropoietin (EPO) fractions differing in sialic acid content, ranging from 9.5 to 13.8 mol mol–1 of EPO, were obtained from baby hamster kidney cell-derived recombinant human EPO by Mono Q column chromatography. The mean pI values of the EPO fractions determined by IEF-gel electrophoresis systematically shifted from 4.11 to 3.31, coinciding with the sialic acid content, without a change in the constitution of asialo N-linked oligosaccharides of each fraction. Although a linear relationship between thein vivo bioactivity and the sialic acid content of the fractionated, samples was observed until 12.1 mol mol–1 of EPO, there was no further increase in their activity over 12.4 mol mol–1 of EPO. On the other hand, an inverse relationship between thein vitro bioactivity and sialic acid content of EPO was observed. Also, we showed that thein vivo bioactivity of some fractions with low sialic acid contents was increased after treatment with 2,6-sialyltransferase, but thein vivo bioactivity of the other fractions with high sialic acid contents was either decreased or not affected.Abbreviations EPO
erythropoietin
- rHuEPO
recombinant human erythropoietin
- hCG
human chorionic gonadotropin
- BHK
baby hamster kidney
- CHO
Chinese hamster ovary
- NeuAc
N-acetyl neuraminic acid
- Gal
galactose
- HRCs
hemolyser-resistant cells
- WST-1
2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium Na
- IEF
isoelectric focusing
- pI
isoelectric point 相似文献
116.
云南姬鼠的蛋白多态性及其遗传分化关系 总被引:3,自引:0,他引:3
本文采用蛋白电泳技术对来源于云南省若干地区的姬鼠属(Apodemus)的3种姬鼠──高山姬鼠(A.chevrieri)8只,中华姬鼠(A.draco)3只和大耳姬鼠(A.latronum)1只,以及作为外群的同科的绒鼠属的大绒鼠(Hapalomysdelalori)3只进行了分析。共检测遗传座位27个,发现21个座位存在多态性。根据蛋白多态的数据对研究对象进行遗传分化关系的探讨,用系统分析软件PHYLIP计算它们之间的分化关系,得到了一棵无根系统树。结果表明,作为外群的大绒鼠明显不同于其它3种姬鼠而聚在最外面。8只高山姬鼠个体汇聚成独立的一支,中华姬鼠的3个个体也聚成一支,但大耳姬鼠却聚在中华姬鼠一支中,因此我们认为大耳姬鼠同中华姬鼠的分化时间可能比较晚近。 相似文献
117.
118.
复合淀粉凝胶电泳同工酶分析 总被引:2,自引:0,他引:2
为了克服水解马铃薯淀粉不易获得的困难,并使“I发片淀粉凝胶电泳同工酶分析”更容易开展,普通的化学试剂马铃薯淀粉(或精制食用马铃薯淀粉)和可溶性淀粉混合物加入适当 剂被用来代替水解马铃薯淀粉制作凝胶。试验结果表明:用8 ̄10%的上述混合淀粉(5:3),添加1%的琼脂粉和2 ̄4%的蔗糖,所制成的“复合淀粉凝胶”可以很好地被切片,并成功地对许多不同类群的植物材料的PGM、PGI、MDH、AAT、SKDH 相似文献
119.
Chieko Takaya Ayako Kosaka Kyouko Kohno Toshihisa Kusano Ken-Ichi Nakamura 《Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology》1995,112(4):727-732
Tetrahymena thermophila could still swim after incubation of the cell body at 40°C for 30 min, whereas Tetrahymena pyriformis did not show any motility after the treatment. Turbidity measurements revealed that axonemes of T. pyriformis lost ATP-dependent sliding activity by the heat treatment, whereas those of T. thermophilia still had the activity under the same conditions. In connection with this difference in susceptibility to high temperature, the biochemical characteristics of dyneins were compared between the two species of Tetrahymena. Axonemal dyneins from the two species had significant vanadate-sensitive ATPase activity even after the heat treatment. Native gel electrophoresis and the following two-dimensional electrophoresis showed that the outer arm dynein of T. thermophilia is more stable in maintaining native configuration than that of T. pyriformis against the heat treatment, although both treated dyneins keep three (α, β and γ) subunits. Analysis by peptide mapping demonstrated that β- and γ-subunits of the outer arm dynein are considerably different in amino acid sequences between the two species. These results imply that dynein of T. thermophilia changed their amino acid sequences and biochemical characteristics to adapt to high temperature. 相似文献
120.
A general assay for restriction endonucleases and other DNA-modifying enzymes with plasmid substrates 总被引:2,自引:0,他引:2
I. Barry Vipond Geoffrey S. Baldwin Mark Oram Symon G. Erskine Lois M. Wentzell Mark D. Szczelkun Timothy J. Nobbs Stephen E. Halford 《Molecular biotechnology》1995,4(3):259-268
A procedure for measuring the activities of enzymes that alter the covalent structure of DNA is described. The assay utilizes
covalently closed circles of DNA as the substrate and yields quantitative data on the fraction of this DNA converted to both
open-circle and linear forms. 相似文献