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排序方式: 共有140条查询结果,搜索用时 31 毫秒
1.
Joshua R Edwards Evangelos A Diamantakos Jacob D Peuler Peter C Lamar Walter C Prozialeck 《BMC physiology》2007,7(1):1
Background
Ethidium homodimer is a cell-membrane impermeant nuclear fluorochrome that has been widely used to identify necrotic cells in culture. Here, we describe a novel technique for evaluating necrosis of epithelial cells in the proximal tubule that involves perfusing ethidium homodimer through the intact rat kidney. As a positive control for inducing necrosis, rats were treated with 3.5, 1.75, 0.87 and 0.43 mg/kg mercuric chloride (Hg2+, intraperitoneal), treatments which have previously been shown to rapidly cause dose-dependent necrosis of the proximal tubule. Twenty-four h after the administration of Hg2+, ethidium homodimer (5 μM) was perfused through the intact left kidney while the animal was anesthetized. The kidney was then removed, placed in embedding medium, frozen and cryosectioned at a thickness of 5 μm. Sections were permeabilized with -20°C methanol and then stained with 4',6-diamidino-2-phenylindole (DAPI) to label total nuclei. Total cell number was determined from the DAPI staining in random microscopic fields and the number of necrotic cells in the same field was determined by ethidium homodimer labeling. 相似文献2.
C D Warren J Alroy B Bugge P F Daniel S S Raghavan E H Kolodny J J Lamar R W Jeanloz 《FEBS letters》1986,195(1-2):247-252
High-pressure liquid chromatography analysis of oligosaccharides from placentas allowed the diagnosis of alpha-mannosidosis in three litters of kittens. The chromatography also afforded a detailed comparison of the oligosaccharide pattern and levels in placenta, liver, brain, urine and ocular fluid of the affected animals. In all cases, two series of compounds were observed, with one or two residues of N-acetylglucosamine at the reducing terminus, respectively, and between two and nine mannose residues. This pattern is unlike that of human mannosidosis, and resembles that of ruminants, except that the major oligosaccharide contains three mannose residues instead of two. 相似文献
3.
Codon usage in the vertebrate hemoglobins and its implications 总被引:2,自引:0,他引:2
A study of codon usage in vertebrate hemoglobins revealed an evolutionary
trend toward elevated numbers of CpG codon boundary pairs in mammalian
hemoglobin alpha genes. Selection for CpG codon boundaries countering the
generally observed CpG suppression is strongly suggested by these data.
These observations parallel recently published experimental results that
indicate that constitutive expression of the human alpha-globin gene
appears to be determined by regulatory information encoded within the
structural gene. The possibility is raised that, in the absence of
selection, CpG decay can be used to date the evolutionary origin of a
mammalian alpha pseudogene from its active alpha gene.
相似文献
4.
Evolution of crystallins: expression of lens-specific proteins in the blind mammals mole (Talpa europaea) and mole rat (Spalax ehrenbergi) 总被引:3,自引:0,他引:3
Quax-Jeuken Y; Bruisten S; Bloemendal H; de Jong WW; Nevo E 《Molecular biology and evolution》1985,2(4):279-288
The mole (Talpa europaea; Insectivora) and the mole rat (Spalax ehrenbergi;
Rodentia) both have degenerated eyes as a convergent adaptation to
subterranean life. The rudimentary eye lenses of these blind mammals no
longer function in a visual process. The crystallin genes, which display a
lens-specific expression pattern, were studied in these blind mammals and
in related species with normal eyes by hybridizing their genomic DNAs with
probes obtained from cDNA clones for alpha A-, alpha B-, and beta
Bp-crystallins from calf and gamma 3- crystallin from the rat. For all
crystallin genes examined, the hybridization signals of mole and mole rat
genomic DNA were comparable, respectively, with those of shrew and of rat
and mouse, normal-vision representatives of the orders Insectivora and
Rodentia. The expression of the crystallins at the protein level was tested
by using antiserum specific for alpha-crystallin in immunofluorescence
reactions on lens sections of mole and mole rat eyes and by using antisera
against the beta- and gamma-crystallins on sections of the mole eye. All
antisera gave positive fluorescence reactions exclusively with lens tissue
of these blind mammals, indicating that the crystallins are still normally
expressed despite the fact that these lenses have had no function in a
visual process in these mammals for at least many million years. These
findings apparently imply that some unknown selective advantage has
conserved the crystallin genes and their expression after the loss of
normal function of the lenses.
相似文献
5.
Eleven flavonol glycosides and two anthocyanins, only one of which was previously identified, were isolated from the flower petals of okra, Hibiscus esculentus L. On the basis of chromatographic, spectral, and degradative evidence, the following structural assignments were made: quercetin 4′-glucoside, quercetin 7-glucoside, quercetin 5-glucoside, quercetin 3-diglucoside, quercetin 4′-diglucoside, quercetin 3-triglucoside, quercetin 5-rhamnoglucoside, gossypetin 8-glucoside, gossypetin 8-rhamnoglucoside, gossypetin 3-glucosido-8-rhamnoglucoside, cyanidin 4′-glucoside, and cyanidin 3-glucosido-4′ glucoside. Some evidence was obtained of a pentahy-droxy, monomethoxy-flavone glycoside. The total flavonoid content in the red portion of the petal was 0.48% of fresh weight; that in the white portion was 2.51%. The two anthocyanins comprised 28.5% of the flavonoid content of the red flower but only a trace of the content of the white. 相似文献
6.
Modulation of ζ-Protein Kinase C by Cyclic AMP in PC12 Cells Occurs Through Phosphorylation by Protein Kinase A 总被引:1,自引:1,他引:0
Marie W. Wooten M. Lamar Seibenhener Laura H. Matthews Guisheng Zhou Elaine S. Coleman 《Journal of neurochemistry》1996,67(3):1023-1031
Abstract: Although cyclic AMP (cAMP) has been reported to cross talk with the protein kinase C (PKC) system, effects of elevated intracellular cAMP on the activities of specific PKC isoforms have not been studied. We report findings from a permeabilized cell assay that was used to examine changes in the activity of the atypical PKC isoforms brought about by exposure of PC12 cells to agents that elevate intracellular cAMP. We found that increases in intracellular cAMP led to rapid stimulation of atypical PKC activity, 40–70% above control, for a sustained period of time, a response that occurred independent of the phorbol 12-myristate 13-acetate (PMA)-sensitive PKC isoforms. Changes in intracellular cAMP levels resulted in a dose-dependent redistribution of ζ-PKC to the cytoplasm with a concomitant increase in the phosphorylation state of the enzyme. Incubation of purified ζ-PKC with increasing concentrations of PKA likewise caused a twofold increase in the phosphorylation state of ζ-PKC. In contrast to the positive effect that PKA-mediated phosphorylation had on the activity of ζ-PKC, the enzyme displayed reduced binding to ras when phosphorylated. Taken together, these findings are consistent with the hypothesis that protein phosphorylation of PKC acts as a positive effector of its enzyme activity and may serve as a negative modulator for interaction with other proteins. 相似文献
7.
Polymerization of pentachlorophenol and ferulic acid by fungal extracellular lignin-degrading enzymes. 总被引:4,自引:0,他引:4
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High-molecular-weight polymers were produced by a crude concentrated supernatant from ligninolytic Phanerochaete chrysosporium cultures in a reaction mixture containing pentachlorophenol and a humic acid precursor (ferulic acid) in the presence of a detergent and H2O2. Pure manganese peroxidase, lignin peroxidase, and laccase were also shown to catalyze the reaction. 相似文献
8.
In Drosophila pseudoobscura, the amylase (Amy) multigene family is
contained within a series of inversions, or gene arrangements, on the third
chromosome. The Standard (ST), Santa Cruz (SC), and Tree Line (TL)
inversions are central to the phylogeny of arrangements, and have clusters
of other arrangements derived from them. The gene arrangements belonging to
each of these three clusters have a characteristic number of Amy genes,
ranging from three in ST to two in SC to one in TL. This distribution
pattern can reflect a history of either duplications or deletions, although
the data available in the past did not permit a decision between these
alternatives. We provide unambiguous evidence that three Amy genes were
present before the divergence of the ST, SC, and TL arrangements. Thus, the
current status of the Amy multigene family is the result of deletions in
the TL and SC arrangements, which created three new pseudogenes: TL
Amy2-psi, TL Amy3-psi, and SC Amy3- psi. Analysis of pseudogene sequences
revealed that, in the SC and ST arrangements, pseudogene evolution has been
retarded, most likely due to the homogenization effect of gene conversion.
Finally, by determining the original copy number, we have reconstructed the
evolutionary history of the Amy multigene family and linked it with the
evolution of the central gene arrangements.
相似文献
9.
Degradation of 4,4'-dichlorobiphenyl, 3,3',4,4'-tetrachlorobiphenyl, and 2,2',4,4',5,5'-hexachlorobiphenyl by the white rot fungus Phanerochaete chrysosporium. 总被引:2,自引:1,他引:1
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The white rot fungus Phanerochaete chrysosporium has demonstrated abilities to degrade many xenobiotic chemicals. In this study, the degradation of three model polychlorinated biphenyl (PCB) congeners (4,4'-dichlorobiphenyl [DCB], 3,3',4,4'-tetrachlorobiphenyl, and 2,2',4,4',5,5'-hexachlorobiphenyl) by P. chrysosporium in liquid culture was examined. After 28 days of incubation, 14C partitioning analysis indicated extensive degradation of DCB, including 11% mineralization. In contrast, there was negligible mineralization of the tetrachloro- or hexachlorobiphenyl and little evidence for any significant metabolism. With all of the model PCBs, a large fraction of the 14C was determined to be biomass bound. Results from a time course study done with 4,4'-[14C]DCB to examine 14C partitioning dynamics indicated that the biomass-bound 14C was likely attributable to nonspecific adsorption of the PCBs to the fungal hyphae. In a subsequent isotope trapping experiment, 4-chlorobenzoic acid and 4-chlorobenzyl alcohol were identified as metabolites produced from 4,4'-[14C]DCB. To the best of our knowledge, this the first report describing intermediates formed by P. chrysosporium during PCB degradation. Results from these experiments suggested similarities between P. chrysosporium and bacterial systems in terms of effects of congener chlorination degree and pattern on PCB metabolism and intermediates characteristic of the PCB degradation process. 相似文献
10.
The alpha-amylase (Amy) multigene family in Drosophila pseudoobscura is
located on the third chromosome, which is polymorphic for more than 40
inverted gene arrangements. The number of copies in this family ranges from
one to three, depending on the arrangement in question. A previous study of
the three Amy genes from the Standard (ST) arrangement suggested either
that duplicated copies (Amy2 and Amy3) are functionally constrained or that
they are undergoing gene conversion with Amy1. In order to elucidate
further the pattern of molecular evolution in this family, we cloned and
sequenced four additional Amy genes, two from the Santa Cruz (SC) and two
from the Chiricahua (CH) gene arrangement. Of the two alternatives, only
the hypothesis of gene conversion is supported by the sequence analysis.
The homogenization effect of gene conversion has been strongest in SC,
whose copies differ by only two nucleotides, less noticeable in ST, and
negligible in the CH. Furthermore, the action of gene conversion is
apparently localized, occurring only in the coding region. Interestingly,
these results concur with the findings of other workers for the duplicated
Amy genes in the Drosophila melanogaster group. Thus, the occurrence of
gene conversion in the Amy multigene family seems to be a common feature in
the Drosophila species studied so far.
相似文献