Grosjean H., Keith G., Droogmans L.
Detection and quantification of modified nucleotides in RNA using thin-layer chromatography Book Section
Dans: Gott, J. M. (Ed.): RNA Interference, Editing, and Modification: Methods and Protocols, vol. 265, p. 357-91, Springer Protocols, Humana Press, 2004.
Résumé | BibTeX | Étiquettes: &, 5', acids, and, Base, Chromatography, Composition, Deoxyribonucleotides/chemistry/isolation, DNA/chemistry/genetics/isolation, Endoribonucleases, Gov't, Indicators, Isotope, KEITH, Labeling/methods, Layer/methods, Non-U.S., Nucleic, Oligoribonucleotides/chemistry/isolation, Peptide, purification, Radioisotopes, Reagents, Regions/chemistry, Ribonucleotides/*analysis, RNA/*genetics/isolation, Support, Thin, Untranslated
@incollection{,
title = {Detection and quantification of modified nucleotides in RNA using thin-layer chromatography},
author = { H. Grosjean and G. Keith and L. Droogmans},
editor = { J.M. Gott},
year = {2004},
date = {2004-01-01},
booktitle = {RNA Interference, Editing, and Modification: Methods and Protocols},
volume = {265},
pages = {357-91},
publisher = {Springer Protocols, Humana Press},
series = {Methods in Molecular Biology},
abstract = {Identification of a modified nucleotide and its localization within an RNA molecule is a difficult task. Only direct sequencing of purified RNA molecules and high-performance liquid chromatography mass spectrometry analysis of purified RNA fragments allow determination of both the type and location of a given modified nucleotide within an RNA of 50-150 nt in length. The objective of this chapter is to describe in detail a few simple procedures that we have found particularly suited for the detection, localization, and quantification of modified nucleotides within an RNA of known sequence. The methods can also be used to reveal the enzymatic activity of a particular RNA-modifying enzyme in vitro or in vivo. The procedures are based on the use of radiolabeled RNA (with [32P], [14C], or [3H]) or [32P]-postlabeled oligonucleotides and two-dimensional thin-layer chromatography of labeled nucleotides on cellulose plates. This chapter provides useful maps of the migration characteristics of 70 modified nucleotides on thin-layer cellulose plates.},
keywords = {&, 5', acids, and, Base, Chromatography, Composition, Deoxyribonucleotides/chemistry/isolation, DNA/chemistry/genetics/isolation, Endoribonucleases, Gov't, Indicators, Isotope, KEITH, Labeling/methods, Layer/methods, Non-U.S., Nucleic, Oligoribonucleotides/chemistry/isolation, Peptide, purification, Radioisotopes, Reagents, Regions/chemistry, Ribonucleotides/*analysis, RNA/*genetics/isolation, Support, Thin, Untranslated},
pubstate = {published},
tppubtype = {incollection}
}
Zukiel R., Nowak S., Barciszewska A. M., Gawronska I., Keith G., Barciszewska M. Z.
A simple epigenetic method for the diagnosis and classification of brain tumors Article de journal
Dans: Mol Cancer Res, vol. 2, no. 3, p. 196-202, 2004, (1541-7786 Journal Article).
Résumé | BibTeX | Étiquettes: *DNA, *Epigenesis, 5-Methylcytosine/*analysis, Adult, Aged, and, Brain, Chromatography, DNA, Female, Genetic, Gov't, Human, KEITH, Layer, Male, Methylation, Middle, Neoplasm/*chemistry/*metabolism, Neoplasms/*classification/*diagnosis/genetics/pathology, Non-U.S., Oxidative, oxygen, Reactive, Sensitivity, Species/metabolism, Specificity, Stress, Support, Thin
@article{,
title = {A simple epigenetic method for the diagnosis and classification of brain tumors},
author = { R. Zukiel and S. Nowak and A. M. Barciszewska and I. Gawronska and G. Keith and M. Z. Barciszewska},
year = {2004},
date = {2004-01-01},
journal = {Mol Cancer Res},
volume = {2},
number = {3},
pages = {196-202},
abstract = {The new, simple, and reliable method for the diagnosis of brain tumors is described. It is based on a TLC quantitative determination of 5-methylcytosine (m(5)C) in relation to its damage products of DNA from tumor tissue. Currently, there is evidence that oxidative stress through reactive oxygen species (ROS) plays an important role in the etiology and progression of several human diseases. Oxidative damage of DNA, lipids, and proteins is deleterious for the cell. m(5)C, along with other basic components of DNA, is the target for ROS, which results in the appearance of new modified nucleic acid bases. If so, m(5)C residue constitutes a mutational hotspot position, whether it occurs within a nucleotide sequence of a structural gene or a regulatory region. Here, we show the results of the analysis of 82 DNA samples taken from brain tumor tissues. DNA was isolated and hydrolyzed into nucleotides, which, after labeling with [gamma-(32)P]ATP, were separated on TLC. Chromatograms were evaluated using PhosphorImager and the amounts of 5-methyldeoxycytosine (m(5)dC) were calculated as a ratio (R) of m(5)dC to m(5)dC + deoxycytosine + deoxythymidine spot intensities. The R value could not only be a good diagnostic marker for brain tumors but also a factor differentiating low-grade and high-grade gliomas. Therefore, DNA methylation pattern might be a useful tool to give a primary diagnosis of a brain tumor or as a marker for the early detection of the relapse of the disease. This method has several advantages over those existing nowadays.},
note = {1541-7786
Journal Article},
keywords = {*DNA, *Epigenesis, 5-Methylcytosine/*analysis, Adult, Aged, and, Brain, Chromatography, DNA, Female, Genetic, Gov't, Human, KEITH, Layer, Male, Methylation, Middle, Neoplasm/*chemistry/*metabolism, Neoplasms/*classification/*diagnosis/genetics/pathology, Non-U.S., Oxidative, oxygen, Reactive, Sensitivity, Species/metabolism, Specificity, Stress, Support, Thin},
pubstate = {published},
tppubtype = {article}
}
Salah R. Ben, Zouari N., Reinbolt J., Mejdoub H.
Purification of turkey pancreatic phospholipase A2 Article de journal
Dans: Biosci Biotechnol Biochem, vol. 67, no. 10, p. 2139-44, 2003, (0916-8451 Journal Article).
Résumé | BibTeX | Étiquettes: *Turkeys, &, A/*isolation, Acid, acids, Amino, Ammonium, and, Animals, Bile, Calcium, Chromatography, Concentration, Data, Hydrogen-Ion, Molecular, Pancreas/*enzymology, Phospholipases, purification, Salts, Sequence, Sulfate, Temperature, Weight
@article{,
title = {Purification of turkey pancreatic phospholipase A2},
author = { R. Ben Salah and N. Zouari and J. Reinbolt and H. Mejdoub},
year = {2003},
date = {2003-01-01},
journal = {Biosci Biotechnol Biochem},
volume = {67},
number = {10},
pages = {2139-44},
abstract = {Turkey pancreatic phospholipase (TPP) has been purified from delipidated pancreases. The purification included ammonium sulfate fractionation, acidic (pH 5) treatment, followed by sequencial column chromatographies on DEAE-cellulose, Sephadex G-75, and reverse phase high pressure liquid chromatography. The purified enzyme was found to be a monomeric protein with molecular mass of 14 kDa. The optimal activity was measured at pH 8 and 37 degrees C using egg yolk emulsion as substrate. Our results show that the enzyme (TPP) was not stable for 1 h at 60 degrees C, and that bile salt and Ca2+ were required for the expression of the purified enzyme. The sequence of the N-terminal amino acids of the purified enzyme shows a very close similarity between TPP and all other known pancreatic phospholipases.},
note = {0916-8451
Journal Article},
keywords = {*Turkeys, &, A/*isolation, Acid, acids, Amino, Ammonium, and, Animals, Bile, Calcium, Chromatography, Concentration, Data, Hydrogen-Ion, Molecular, Pancreas/*enzymology, Phospholipases, purification, Salts, Sequence, Sulfate, Temperature, Weight},
pubstate = {published},
tppubtype = {article}
}
Popiela A., Gabrys M. S., Rabczynski J., Panszczyk M., Keith G., Baranowski W.
[Estimation of DNA methylation level in endometrial cancer tissues] Article de journal
Dans: Ginekol Pol, vol. 73, no. 11, p. 966-9, 2002, (0017-0011 Journal Article).
Résumé | BibTeX | Étiquettes: *DNA, 80, Abstract, Adenocarcinoma/chemistry/*genetics, Aged, and, Case-Control, Endometrial, English, Expression, Female, Gene, Human, Hyperplasia/genetics, Methylation, Middle, Neoplasms/chemistry/*genetics, Neoplastic, over, Regulation, Studies
@article{,
title = {[Estimation of DNA methylation level in endometrial cancer tissues]},
author = { A. Popiela and M. S. Gabrys and J. Rabczynski and M. Panszczyk and G. Keith and W. Baranowski},
year = {2002},
date = {2002-01-01},
journal = {Ginekol Pol},
volume = {73},
number = {11},
pages = {966-9},
abstract = {OBJECTIVES: A level of DNA methylation plays an important role in regulation of cellular gene's expression. Estimation of DNA methylation level in endometrial neoplastic tissues compared to normal endometrial samples was the aim of this study. DESIGN: It was to be shown, that changes in methylation rate in promotory regions could lead to carcinogenesis in particular cell. Authors describe an analysis of DNA methylation level in endometrial cancer tissues compared to DNA methylation level in normal or hyperplastic endometrium. MATERIAL AND METHODS: Endometrial samples from 88 women were collected. 56 of them were classified as adenocarcinoma, 20 as hyperplastic changes, 12 as normal endometrium-control group. DNA was isolated from tissues and than prepared to pm5dC and pdC. Than we performed a statistical analysis of results. RESULTS: The median DNA methylation level was significantly higher in neoplastic tissues than in normal endometrium. There was no difference between DNA methylation level between normal endometrium and hyperplastic changes. CONCLUSIONS: Authors conclude that neoplastic endometrial tissues show high DNA methylation rate compared to normal or hyperplastic endometrium.},
note = {0017-0011
Journal Article},
keywords = {*DNA, 80, Abstract, Adenocarcinoma/chemistry/*genetics, Aged, and, Case-Control, Endometrial, English, Expression, Female, Gene, Human, Hyperplasia/genetics, Methylation, Middle, Neoplasms/chemistry/*genetics, Neoplastic, over, Regulation, Studies},
pubstate = {published},
tppubtype = {article}
}
Popiela A., Gabrys M. S., Rabczynski J., Panszczyk M., Keith G., Baranowski W.
[Estimation of DNA methylation level in nonendometrial uterus malignancies] Article de journal
Dans: Ginekol Pol, vol. 73, no. 11, p. 962-5, 2002, (0017-0011 Journal Article).
Résumé | BibTeX | Étiquettes: *DNA, 80, Abstract, Age, Aged, and, Case-Control, English, Expression, Factors, Female, Gene, Human, Mesodermal/genetics/*metabolism, Methylation, Middle, Mixed, Neoplasms/genetics/*metabolism, Neoplastic, over, Regulation, silencing, Studies, tumor, Uterine
@article{,
title = {[Estimation of DNA methylation level in nonendometrial uterus malignancies]},
author = { A. Popiela and M. S. Gabrys and J. Rabczynski and M. Panszczyk and G. Keith and W. Baranowski},
year = {2002},
date = {2002-01-01},
journal = {Ginekol Pol},
volume = {73},
number = {11},
pages = {962-5},
abstract = {OBJECTIVES: Rebuilding of genome structure leads to many pathological states including neoplastic malignancies. Rebuilding often occurs as a process caused by disturbances in gene silencing mechanism. DNA methylation pattern is one of the most important mechanisms connected to gene's silencing. Estimation of DNA methylation level in nonendometrial uterine neoplastic tissues compared to normal endometrial samples was the aim of this study. DESIGN: It was to be shown, that changes in methylation rate in promotory regions could lead to carcinogenesis in particular cell. Authors describe an analysis of DNA methylation level in nonendometrial neoplastic uterine tissues compared to DNA methylation level in normal endometrium. MATERIALS AND METHODS: Tissue samples from 9 women with tumor mixtus mesodermalis were collected. 12 samples were normal endometrium-control group. DNA was isolated from tissues and than we performed an estimation of DNA methylation levels. Than we performed a statistical analysis of results. RESULTS: The median DNA methylation level was significantly higher in neoplastic tissues than in normal endometrium. CONCLUSIONS: Authors conclude, that DNA methylation level is higher in neoplastic tissues, but does not correlate with clinical stage of the disease.},
note = {0017-0011
Journal Article},
keywords = {*DNA, 80, Abstract, Age, Aged, and, Case-Control, English, Expression, Factors, Female, Gene, Human, Mesodermal/genetics/*metabolism, Methylation, Middle, Mixed, Neoplasms/genetics/*metabolism, Neoplastic, over, Regulation, silencing, Studies, tumor, Uterine},
pubstate = {published},
tppubtype = {article}
}
Postawski K., Olech-Fudali E., Jakowicki J. A., Korobowicz E., Keith G., Baranowski W.
[Overall genomic DNA methylation in relation to estrogen] Article de journal
Dans: Ginekol Pol, vol. 71, no. 9, p. 1206-11, 2000, (0017-0011 Journal Article).
Résumé | BibTeX | Étiquettes: 80, Abstract, Aged, and, Biopsy, DNA, English, Estrogen/*metabolism, Female, Gov't, Human, Methylases/metabolism, Methylation, Middle, modification, Neoplasms/*metabolism/*pathology, Non-U.S., over, Progesterone/*metabolism, Receptors, Support, Uterine, Uterus/*metabolism/pathology
@article{,
title = {[Overall genomic DNA methylation in relation to estrogen]},
author = { K. Postawski and E. Olech-Fudali and J. A. Jakowicki and E. Korobowicz and G. Keith and W. Baranowski},
year = {2000},
date = {2000-01-01},
journal = {Ginekol Pol},
volume = {71},
number = {9},
pages = {1206-11},
abstract = {Overall genomic DNA methylation was analyzed using enzymatic digestion into nucleotides, 32P postlabeling, two-dimensional thin-layer chromatography on cellulose plates and phosphobioimaging quantitation, in relation to immunohistochemically measured estrogen (ER) and progesterone receptor (PR) status of 15 uterine cancers. Mean 5-methyldeoxycytosine (m5dC) content did not differ between ER-positive and ER-negative neoplasms. Highest values of m5dC were noted both in ER-negative and ER-positive tumors. Additionally, there was no low DNA methylation in ER negative uterine cancer tissues. Decrease of the overall genomic DNA methylation could be related to the increase of ER/PR ratio, however it was not significant in our investigation. The potential role of steroid receptors status in uterine cancer tissue is discussed.},
note = {0017-0011
Journal Article},
keywords = {80, Abstract, Aged, and, Biopsy, DNA, English, Estrogen/*metabolism, Female, Gov't, Human, Methylases/metabolism, Methylation, Middle, modification, Neoplasms/*metabolism/*pathology, Non-U.S., over, Progesterone/*metabolism, Receptors, Support, Uterine, Uterus/*metabolism/pathology},
pubstate = {published},
tppubtype = {article}
}
Keith G., Dirheimer G.
Postlabeling: a sensitive method for studying DNA adducts and their role in carcinogenesis Article de journal
Dans: Curr Opin Biotechnol, vol. 6, no. 1, p. 3-11, 1995, (0958-1669 Journal Article Review Review, Academic).
Résumé | BibTeX | Étiquettes: *Cell, *Genome, Adducts/*analysis, and, Animals, Base, Cell, Conditions/genetics/pathology, Data, Dilution, Division, DNA, Genetic, Gov't, Human, Models, Molecular, Mutagenesis, Neoplasms/*genetics/pathology, Neoplastic, Non-U.S., Phosphorus, Precancerous, Radioisotope, Radioisotopes, Sensitivity, Sequence, Specificity, Support, Technique, Transformation, Xenobiotics
@article{,
title = {Postlabeling: a sensitive method for studying DNA adducts and their role in carcinogenesis},
author = { G. Keith and G. Dirheimer},
year = {1995},
date = {1995-01-01},
journal = {Curr Opin Biotechnol},
volume = {6},
number = {1},
pages = {3-11},
abstract = {The covalent binding of xenobiotics to DNA is an important trigger of the multistage process that leads to carcinogenesis. 32P-postlabeling represents a highly sensitive method for biomonitoring exposure to genotoxic agents and for cancer risk assessment; it is capable of detecting less than one DNA adduct per human genome. Recent improvements to the technique have shown that the resistance of adducted DNA to enzyme digestion may lead to an overestimation of the number of different adducts present in a sample.},
note = {0958-1669
Journal Article
Review
Review, Academic},
keywords = {*Cell, *Genome, Adducts/*analysis, and, Animals, Base, Cell, Conditions/genetics/pathology, Data, Dilution, Division, DNA, Genetic, Gov't, Human, Models, Molecular, Mutagenesis, Neoplasms/*genetics/pathology, Neoplastic, Non-U.S., Phosphorus, Precancerous, Radioisotope, Radioisotopes, Sensitivity, Sequence, Specificity, Support, Technique, Transformation, Xenobiotics},
pubstate = {published},
tppubtype = {article}
}