Application of proton NMR spectroscopy to measurement of whole-body RNA degradation rates: effects of surgical stress in human patients
The urinary catabolites, N 2, N 2-dimethylguanosine (DMG), pseudouridine (PSU) and 7-methylguanine (m 7-Gua) are formed from post-transcriptional methylation of RNA bases and are not reincorporated into RNA upon its degradation. Their quantitative urinary excretion may be used to determine rates of...
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Published in | Clinica chimica acta Vol. 252; no. 2; pp. 123 - 135 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Shannon
Elsevier B.V
30.08.1996
Elsevier |
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Abstract | The urinary catabolites,
N
2,
N
2-dimethylguanosine (DMG), pseudouridine (PSU) and 7-methylguanine (m
7-Gua) are formed from post-transcriptional methylation of RNA bases and are not reincorporated into RNA upon its degradation. Their quantitative urinary excretion may be used to determine rates of whole body degradation of individual RNA species since DMG occurs exclusively in tRNA, PSU occurs in rRNA and tRNA and m
7-Gua occurs in all RNA species. Conventional HPLC analysis has several drawbacks since pre-analytical steps may involve selective losses and, under certain conditions, other urinary analytes may co-elute. In the present paper, we report analysis of these compounds by high-field
1H-nuclear magnetic resonance (
1H-NMR) spectroscopy. Urinary concentrations of these metabolites were found to be in agreement with previously published HPLC and ELISA determinations. However, NMR analysis required minimal sample preparation (other than lyophilisation and reconstitution) and was capable of the simultaneous determination of other relevant analytes such as creatinine. This technique was therefore applied to urine samples from patients who had undergone surgical stress and insulin-like growth factor-I (IGF-I) therapy. Surgical stress increased the excretion of DMG and m
7-Gua. Degradation rates for tRNA and mRNA were also higher in surgically stressed subjects when compared with controls but degradation rates of rRNA decreased by approx. 30%. However, injection of IGF-I (40 μg/kg s.c.) had no significant effect on the excretion of these nucleosides. These data indicated that IGF-I therapy has no marked effects on RNA turnover following trauma. We suggest that this technique can be applied to study of RNA metabolism in any surgical or medical condition. Furthermore, since only 0.6 ml of urine is required, studies in neonates seem to be feasible. |
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AbstractList | The urinary catabolites, N2,N2-dimethylguanosine (DMG), pseudouridine (PSU) and 7-methylguanine (m7-Gua) are formed from post-transcriptional methylation of RNA bases and are not reincorporated into RNA upon its degradation. Their quantitative urinary excretion may be used to determine rates of whole body degradation of individual RNA species since DMG occurs exclusively in tRNA, PSU occurs in rRNA and tRNA and m7-Gua occurs in all RNA species. Conventional HPLC analysis has several drawbacks since pre-analytical steps may involve selective losses and, under certain conditions, other urinary analytes may co-elute. In the present paper, we report analysis of these compounds by high-field 1H-nuclear magnetic resonance (1H-NMR) spectroscopy. Urinary concentrations of these metabolites were found to be in agreement with previously published HPLC and ELISA determinations. However, NMR analysis required minimal sample preparation (other than lyophilisation and reconstitution) and was capable of the simultaneous determination of other relevant analytes such as creatinine. This technique was therefore applied to urine samples from patients who had undergone surgical stress and insulin-like growth factor-1 (IGF-I) therapy. Surgical stress increased the excretion of DMG and m7-Gua. Degradation rates for tRNA and mRNA were also higher in surgically stressed subjects when compared with controls but degradation rates of rRNA decreased by approx. 30%. However, injection of IGF-I (40 micrograms/kg s.c.) had no significant effect on the excretion of these nucleosides. These data indicated that IGF-I therapy has no marked effects on RNA turnover following trauma. We suggest that this technique can be applied to study of RNA metabolism in any surgical or medical condition. Furthermore, since only 0.6 ml of urine is required, studies in neonates seem to be feasible. The urinary catabolites, N 2, N 2-dimethylguanosine (DMG), pseudouridine (PSU) and 7-methylguanine (m 7-Gua) are formed from post-transcriptional methylation of RNA bases and are not reincorporated into RNA upon its degradation. Their quantitative urinary excretion may be used to determine rates of whole body degradation of individual RNA species since DMG occurs exclusively in tRNA, PSU occurs in rRNA and tRNA and m 7-Gua occurs in all RNA species. Conventional HPLC analysis has several drawbacks since pre-analytical steps may involve selective losses and, under certain conditions, other urinary analytes may co-elute. In the present paper, we report analysis of these compounds by high-field 1H-nuclear magnetic resonance ( 1H-NMR) spectroscopy. Urinary concentrations of these metabolites were found to be in agreement with previously published HPLC and ELISA determinations. However, NMR analysis required minimal sample preparation (other than lyophilisation and reconstitution) and was capable of the simultaneous determination of other relevant analytes such as creatinine. This technique was therefore applied to urine samples from patients who had undergone surgical stress and insulin-like growth factor-I (IGF-I) therapy. Surgical stress increased the excretion of DMG and m 7-Gua. Degradation rates for tRNA and mRNA were also higher in surgically stressed subjects when compared with controls but degradation rates of rRNA decreased by approx. 30%. However, injection of IGF-I (40 μg/kg s.c.) had no significant effect on the excretion of these nucleosides. These data indicated that IGF-I therapy has no marked effects on RNA turnover following trauma. We suggest that this technique can be applied to study of RNA metabolism in any surgical or medical condition. Furthermore, since only 0.6 ml of urine is required, studies in neonates seem to be feasible. |
Author | Marway, Jaspaul S. Peters, Timothy J. Ross, Richard Grimble, George K. Preedy, Victor R. Miell, John P. Gibbons, William A. Bonner, Adrian B. Anderson, Graeme J. |
Author_xml | – sequence: 1 givenname: Jaspaul S. surname: Marway fullname: Marway, Jaspaul S. organization: Tissue Pathology Unit, Roehampton Institute London, West Hill, London, SW15 3SN, UK – sequence: 2 givenname: Graeme J. surname: Anderson fullname: Anderson, Graeme J. organization: Department of Chemistry, Manchester Metropolitan University, Chester Street, Manchester M1 5GD, UK – sequence: 3 givenname: John P. surname: Miell fullname: Miell, John P. organization: Department of Medicine, King's College School of Medicine and Dentistry, Bessemer Road, London, SE5 9PJ, UK – sequence: 4 givenname: Richard surname: Ross fullname: Ross, Richard organization: Department of Medicine, King's College School of Medicine and Dentistry, Bessemer Road, London, SE5 9PJ, UK – sequence: 5 givenname: George K. surname: Grimble fullname: Grimble, George K. organization: Tissue Pathology Unit, Roehampton Institute London, West Hill, London, SW15 3SN, UK – sequence: 6 givenname: Adrian B. surname: Bonner fullname: Bonner, Adrian B. organization: Tissue Pathology Unit, Roehampton Institute London, West Hill, London, SW15 3SN, UK – sequence: 7 givenname: William A. surname: Gibbons fullname: Gibbons, William A. organization: Department of Pharmaceutical Chemistry, School of Pharmacy, London, UK – sequence: 8 givenname: Timothy J. surname: Peters fullname: Peters, Timothy J. organization: Department of Clinical Biochemistry, King's College School of Medicine and Dentistry, Bessemer Road, London, SE5 9PJ, UK – sequence: 9 givenname: Victor R. surname: Preedy fullname: Preedy, Victor R. organization: Department of Clinical Biochemistry, King's College School of Medicine and Dentistry, Bessemer Road, London, SE5 9PJ, UK |
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Cites_doi | 10.1016/0002-9378(83)90962-6 10.1042/cs0800393 10.1002/1097-0142(197508)36:2<390::AID-CNCR2820360214>3.0.CO;2-C 10.1042/cs0720563 10.1182/blood.V61.2.291.291 10.1016/S0149-2918(05)80001-3 10.1016/S0021-9673(01)89352-3 10.1177/0148607186010006578 10.1016/0026-0495(84)90046-5 10.1042/cs0710367 10.1016/0167-4889(86)90177-1 10.1111/j.1365-2265.1992.tb01486.x 10.1038/241204a0 10.1016/0167-4781(82)90128-2 10.1042/bj2670325 10.1042/bj0241244 10.1042/bj1780139 10.1016/0009-8981(93)90181-3 10.1093/oxfordjournals.alcalc.a045158 10.1016/0261-5614(86)90020-8 10.1016/0009-8981(92)90087-7 10.1136/bmj.289.6445.584 10.1111/j.1365-2265.1991.tb03495.x |
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Keywords | Urine m 7-Gua, 7-methylguanine IGF-I DMG, N 2, N 2-dimethylguanosine Surgical stress Proton nuclear magnetic resonance RNA catabolites PSU, pseudouridine Human Biological fluid Investigation method RNA Surgery Exploration NMR spectrometry Degradation product Trauma |
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N
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N
2-dimethylguanosine (DMG), pseudouridine (PSU) and 7-methylguanine (m
7-Gua) are formed from post-transcriptional methylation... The urinary catabolites, N2,N2-dimethylguanosine (DMG), pseudouridine (PSU) and 7-methylguanine (m7-Gua) are formed from post-transcriptional methylation of... |
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SubjectTerms | Adult Aged Biological and medical sciences Humans Hydrolysis IGF-I Insulin-Like Growth Factor I - therapeutic use Magnetic Resonance Spectroscopy Male Medical sciences Middle Aged Miscellaneous Proton nuclear magnetic resonance Protons Recombinant Proteins - therapeutic use RNA - metabolism RNA catabolites RNA Processing, Post-Transcriptional Surgery (general aspects). Transplantations, organ and tissue grafts. Graft diseases Surgical Procedures, Operative - adverse effects Surgical stress Urine |
Title | Application of proton NMR spectroscopy to measurement of whole-body RNA degradation rates: effects of surgical stress in human patients |
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