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排序方式: 共有174条查询结果,搜索用时 15 毫秒
41.
Biomarkers: mining the biofluid proteome 总被引:8,自引:0,他引:8
Veenstra TD Conrads TP Hood BL Avellino AM Ellenbogen RG Morrison RS 《Molecular & cellular proteomics : MCP》2005,4(4):409-418
Proteomics has brought with it the hope of identifying novel biomarkers for diseases such as cancer. This hope is built on the ability of proteomic technologies, such as mass spectrometry (MS), to identify hundreds of proteins in complex biofluids such as plasma and serum. There are many factors that make this research very challenging beginning with the lack of standardization of sample collection and continuing through the entire analytical process. Fortunately the advances made in the characterization of biofluids using proteomic techniques have been rapid and suggest that these mainly discovery driven approaches will lead to the development of highly specific platforms for diagnosing diseases and monitoring responses to different treatments in the near future. 相似文献
42.
Gillette WK Esposito D Frank PH Zhou M Yu LR Jozwik C Zhang X McGowan B Jacobowitz DM Pollard HB Hao T Hill DE Vidal M Conrads TP Veenstra TD Hartley JL 《Molecular & cellular proteomics : MCP》2005,4(11):1647-1652
We have developed a pooled ORF expression technology, POET, that uses recombinational cloning and proteomic methods (two-dimensional gel electrophoresis and mass spectrometry) to identify ORFs that when expressed are likely to yield high levels of soluble, purified protein. Because the method works on pools of ORFs, the procedures needed to subclone, express, purify, and assay protein expression for hundreds of clones are greatly simplified. Small scale expression and purification of 12 positive clones identified by POET from a pool of 688 Caenorhabditis elegans ORFs expressed in Escherichia coli yielded on average 6 times as much protein as 12 negative clones. Larger scale expression and purification of six of the positive clones yielded 47-374 mg of purified protein/liter. Using POET, pools of ORFs can be constructed, and the pools of the resulting proteins can be analyzed and manipulated to rapidly acquire information about the attributes of hundreds of proteins simultaneously. 相似文献
43.
Yu LR Conrads TP Uo T Issaq HJ Morrison RS Veenstra TD 《Journal of proteome research》2004,3(3):469-477
The new generation of isotope-coded affinity tag (ICAT) reagents have been evaluated by labeling an equimolar amount of bovine serum albumin (BSA) with ICAT-12C9 and ICAT-13C9, combining the mixtures, digesting them with trypsin and analyzing the digestate both by muRPLC-tandem MS and by matrix-assisted laser desorption ionization (MALDI) TOF/TOF MS. The use of 13C in place of 2H resulted in both of the labeled peptides having identical elution characteristics in a reversed-phase separation. This similarity in elution allows ICAT-labeled peptides to be effectively analyzed using a muRPLC-MALDI-MS strategy as well. All of the cysteinyl-containing tryptic peptides from BSA were identified with only a 10% variation in the relative abundance measurements between the light and heavy versions of each peptide. A facile method for the removal of contaminants that arise from the cleaved biotin moiety that otherwise interfere with downstream separations and MS analysis has also been developed. The new ICAT reagents were then applied to the analysis of a cortical neuron proteome sample to identify proteins regulated by the antitumor drug, camptothecin. 相似文献
44.
Blonder J Conrads TP Yu LR Terunuma A Janini GM Issaq HJ Vogel JC Veenstra TD 《Proteomics》2004,4(1):31-45
A simple and rapid method for characterizing hydrophobic integral membrane proteins and its utility for membrane proteomics using microcapillary liquid chromatography coupled on-line with tandem mass spectrometry (microLC-MS/MS) is described. The present technique does not rely on the use of detergents, strong organic acids or cyanogen bromide-mediated proteolysis. A buffered solution of 60% methanol was used to extract, solubilize, and tryptically digest proteins within a preparation of Halobacterium (H.) halobium purple membranes. Analysis of the digested purple membrane proteins by microLC-MS/MS resulted in the identification of all the predicted tryptic peptides of bacteriorhodopsin, including those that are known to be post-translationally modified. In addition, 40 proteins from the purple membrane preparation were also identified, of which 80% are predicted to contain between 1 and 16 transmembrane domains. To evaluate the general applicability of the method, the same extraction, solubilization, and digestion conditions were applied to a plasma membrane fraction prepared from human epidermal sheets. A total of 117 proteins was identified in a single microLC-MS/MS analysis, of which 55% are known to be integral or associated with the plasma membrane. Due to its simplicity, efficiency, and absence of MS interfering compounds, this technique can be used for the characterization of other integral membrane proteins and may be concomitantly applied for the analysis of membrane protein complexes or large-scale proteomic studies of different membrane samples. 相似文献
45.
A major goal of proteomics is to develop methods that enable the systematic characterization of every protein within the cell or particular subcellular proteome using a single analytical platform. Although the equivalent has already been achieved in genomics, reaching this goal in proteomics represents a much greater challenge due to the wide dynamic range of protein expression, numerous post-translational modifications and remarkable physicochemical heterogeneity of proteins. A major analytical challenge has involved developing more effective means for proteome-scale investigations of membrane proteins, whose solubility differs drastically from that of cytoplasmic proteins. Fortunately, rapid progress has increased the ability to characterize this critically important class of proteins on a scale analogous to that of aqueous soluble proteins. 相似文献
46.
Ján Lí?KA Július BRTKO Michal DUBOVICKY Dana MACEJOVá Viktória KISSOVá ?tefan POLáK Eduard UJHáZY 《Experimental Animals》2016,65(1):1-9
The mammary gland is a dynamic organ that undergoes structural and functional changes
associated with growth, reproduction, and post-menopausal regression. The postnatal
transformations of the epithelium and stromal cells of the mammary gland may contribute to
its susceptibility to carcinogenesis. The increased cancer incidence in mammary glands of
humans and similarly of rodents in association with their development is believed to be
partly explained by proliferative activity together with lesser degree of differentiation,
but it is not completely understood how the virgin gland retains its higher susceptibility
to carcinogenesis. During its developmental cycle, the mammary gland displays many of the
properties associated with breast cancer. An early first full-term pregnancy may have a
protective effect. Rodent models are useful for investigating potential breast
carcinogens. The purpose of this review is to help recognizing histological appearance of
the epithelium and the stroma of the normal mammary gland in rats, and throughout its
development in relation to tumorigenic potential. 相似文献
47.
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49.
ELIŠKA ZÁVESKÁ TOMÁŠ FÉR OTAKAR ŠÍDA JANA LEONG‐ŠKORNIČKOVÁ MAMYIL SABU KAROL MARHOLD 《Botanical journal of the Linnean Society. Linnean Society of London》2011,165(4):388-401
The relationships between genome size and the systematic and evolutionary patterns in vascular plants are equivocal, although a close relationship between genome size and evolutionary patterns has been previously reported. However, several studies have also revealed the dynamic nature of genome size evolution and its considerable ‘ups’ and ‘downs’. Thus, in this study, the phylogenetic relationships among three previously revealed genome size groups and among species of the highly polyploid genus Curcuma were evaluated using AFLP. Our results suggest two main lineages within Indian Curcuma reflecting evolution of genome size. The first one includes hexaploids and higher polyploids of the previously recognized genome size group I, and the second one includes mainly hexaploids of genome size groups II and III. Within genome size group I, relationships among species seem to be influenced by reticulate evolution and higher polyploids are likely to be of allopolyploid origin. Reproductive systems in Indian Curcuma vary considerably among ploidy levels and these differences considerably affect morphological and genetic variation. In general, clonally reproducing species are expected to exhibit low genotypic diversity, but, at the same time, species of allopolyploid origin are expected to maintain higher levels of heterozygosity compared with their progenitors. We investigated intra‐populational genetic variability in Curcuma spp. to evaluate whether mode of reproduction or ploidy represent the main factor influencing the degree of genetic diversity. We found that hexaploid species exhibited significantly higher genetic diversity than higher polyploids (9x, 15x). Our results suggest that this genetic diversity pattern is largely influenced by the mode of reproduction, as higher polyploids reproduce exclusively vegetatively, whereas hexaploids reproduce mainly sexually. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 165 , 388–401. 相似文献
50.
The acute toxicity of the aqueous and ethanol extracts of Parkia biglobosa pods against Clarias gariepinus was investigated under laboratory conditions. Agitated behaviours and respiratory distress were also observed during the exposure period. The adverse effects on biochemical parameters were assessed using semi-static bioassays for 28 days. The ethanol extract of P. biglobosa pods was found to be more acutely toxic with a 96 h LC50 value of 13.96 mg l?1 than the aqueous extracts, with a 96 h LC50 value of 19.95 mg l?1 against C. gariepinus. Both extracts induced agitated behaviours and respiratory distress in exposed organisms. The activities of superoxide dismutase (SOD), catalase (CAT) and the concentration of malondialdehyde (MDA) were significantly lower (p < 0.05) in groups of organisms exposed to extracts of P. biglobosa when compared with the control group after 14 days. The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were also significantly (p < 0.05) lower compared with activities of the enzymes in the control group after 28 days. The current study has shown that the introduction of P. biglobosa pods into aquatic ecosystems is acutely toxic to fish and would possibly be to other non-target aquatic organisms especially invertebrates. 相似文献