Chapter 2 Ribonucleosides in Biological Fluids by A High-Resolution Quantitative Rplc-UV Method

Kuo, Kenneth C.; Phan, Dat Tuan; Williams, N.J.; Gehrke, Charles W.

doi:10.1016/s0301-4770(08)61540-7

Cited by 10 publications

(5 citation statements)

References 43 publications

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“…The method described by Kuo et al (15) allowed the separation and quantification of other modified nucleosides in serum using reversedphase HPLC with diode-array measurement. The concentrations of other nucleosides in blood serum are also consistently elevated in patients with malignant proliferative diseases (15,29), but it should be noted that their blood plasma concentrations are 100 times less than that of pseudouridine, and their concentrations also display greater variability, due to nonmetabolic factors (19).…”

Section: Discussionmentioning

confidence: 99%

“…There is a considerable number of publications concerning changes in urinary pseudouridine excretion as a response to malignant growth (6 -14). There is much less information about changes in blood plasma concentrations of pseudouridine, which is 100 times lower than in urine, although it can still be determined precisely with HPLC techniques (15)(16)(17)(18). Advantages of pseudouridine as a marker of RNA turnover rate in comparison with other modified nu- cleosides are its approximately 100-fold greater concentration in blood plasma, and its lesser variability attributable to the renal clearance rate, sex-related differences, genetic influences or environmental conditions (19).…”

Section: Introductionmentioning

confidence: 99%

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Blood Plasma Pseudouridine in Patients with Malignant Proliferative Diseases

Motyl¹,

Traczyk²,

Cieśluk³

et al. 1993

Clinical Chemistry and Laboratory Medicine

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The blood plasma concentration of pseudouridine was estimated in 104 healthy adult subjects, and 108 patients suffering from malignant proliferative diseases. The HPLC method for simultaneous determination of pseudouridine and creatinine was applied.The average physiological concentration of pseudouridine in blood plasma was 2.43 ± 0.97 μπιοί -I" 1 or 29.15 + 7.40 mmol · mol" 1 creatinine. The physiological urinary excretion of pseudouridine was 14.32 ± 5.20 μπιοί -24h" 1 · kg~°· 75 or 19.60 + 5.22 mmol -mol" 1 creatinine. Renal clearance of pseudouridine and endogenous creatinine were 4.04 + 0.99 and 5.50 ± 1.46ml -kg" 075 , respectively. A positive correlation (r = 0.55, P < 0.01) was found between age (in the range 20 -92 years) and blood plasma pseudouridine concentration (μιηοΐ · I" 1 ). By expressing plasma pseudouridine in relation to plasma creatinine, the apparent influence of non-metabolic factors (age, renal insufficiency, blood dilution) on the plasma pseudouridine concentration were largely excluded.Among haematological proliferative diseases the highest values of plasma pseudouridine concentrations were observed in chronic lymphocytic leukaemia (8.19 μπιοί-I" 1 ; 54.9 mmol -mol"" 1 creatinine) and multiple myeloma (7.02 μπιοί · I" 1 ; 52.5 mmol · mol~J creatinine). In multiple myeloma, but not in chronic lymphocytic leukaemia, the plasma pseudouridine concentration depended on the clinical stage. A lower, but still significant response in non^Hodgkin's lymphoma was noted (4.03 μπιοί · I" 1 ; 40.88 mmol · mol" 1 creatinine).A significant increase of the plasma pseudouridine concentration was characteristic of adenocarcinomas of the large intestine, and it occurred in the early stages of malignant growth. In patients with lung cancer the plasma pseudouridine concentration was elevated only in advanced cases with metastases. The increased pseudouridine concentration was evident in all examined cancers of the urogenital system: cancer of the urinary bladder, cancer of the kidney, cancer of the prostate, and cancer of the testis.It is concluded that the determination of pseudouridine in blood plasma, particularly in relation to creatinine, is a valuable biochemical m rker of accelerated turnover rate of nucleic acids associated with neoplastic growth. ') This study was supported by a grant from the Ministry of National Education.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Blood Plasma Pseudouridine in Patients with Malignant Proliferative Diseases

Motyl¹,

Traczyk²,

Cieśluk³

et al. 1993

Clinical Chemistry and Laboratory Medicine

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“…8-oxodG was determined using a solid phase extraction and high performance liquid chromatography (HPLC) method with electrochemical detection as has been described before (26). The method used for measurement of is based on prefractionation of using a boronate gel followed by reversed phase HPLC with UV detection (36,37). Gla was determined using a precolumnderivatization of Gla with o-phthal-dialdehyde followed by reversed HPLC with fluorescence detection as described previously (32).…”

Section: Determination Of 8-oxo-2-deoxyguanosine (8-oxodg) Pseudourimentioning

confidence: 99%

Noninvasive Markers of Oxidative DNA Stress, RNA Degradation and Protein Degradation Are Differentially Correlated With Resting Metabolic Rate and Energy Intake in Children and Adolescents

et al. 2008

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Urinary excreted RNA and DNA catabolites are used as noninvasive markers for metabolic processes: 8-oxo-2'-deoxyguanosine (8-oxodG) potentially represents oxidative stress to DNA/deoxyribonucleotidetriphosphate pool, modified ribonucleoside pseudouridine (psi) originating mainly from degraded rRNA and tRNA reflects RNA turnover. Modified amino acid gamma-carboxyglutamic acid (Gla) stems from degraded proteins reflecting turnover of proteins. Aim of the present study was to investigate (44 healthy males, 3-18 y) how excretion rates of 8-oxodG, psi, and Gla are related to resting metabolic rate and energy intake. Excretion rates of 8-oxodG (pmol/kg/d), psi (micromol/kg/d), and Gla (micromol/kg/d) were significantly correlated with resting metabolic rate (kJ/kg/d): r = 0.108 (p = 0.029), 0.691 and 0.552 (p < 0.0001), respectively. Excretion rates of 8-oxodG, psi, and Gla were also significantly correlated with energy intake (kJ/kg/d): r = 0.108 (p = 0.036), 0.602 and 0.462 (p < 0.0001). 8-oxodG and Gla excretion was significantly correlated with psi excretion: r = 0.174 (p = 0.005) and 0.709 (p < 0.0001). These results indicate close relationships between whole-body RNA and protein degradation and metabolic rate. The relationship between 8-oxodG excretion and metabolic rate, however, is less strong suggesting that factors other than metabolic rate considerably affect the oxidative stress to DNA.

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“…The analytical procedures for measuring levels of the modified RNA catabolites in urine and the evaluation of the method have been described elsewhere [9,10]. The method is based on enrichment of modified ribonucleosides from urine by affinity chromatography using a boronate gel followed by reverse-phase HPLC determination of the ribonucleoside fraction with UV absorption at 254 nm.…”

Section: Determination Of Levels Of Modified Ribonucleosides Creatinmentioning

confidence: 99%

Diurnal variation in the renal excretion of modified RNA catabolites in humans

Topp

Unverzagt²,

Rudloff

et al. 2003

Clinical Science

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Renally excreted modified RNA catabolites [pseudouridine (psi), N (2), N (2)-dimethylguanosine (m(2)(2)G) and N (6)-threoninocarbonyladenosine (t(6)A)] are markers of whole-body rates of degradation of rRNA and tRNA, and are thought to be potential indicators of the resting metabolic rate. To investigate diurnal variations of these RNA catabolites, the amounts of psi, m(2)(2)G and t(6)A excreted were determined by HPLC of the urine from eight healthy male adults collected over 47-h periods, which were subdivided into the morning (06.00 or 09.00 to 12.00 hours), the afternoon (12.00 to 18.00 hours), the evening (18.00 to 24.00 hours) and the night (00.00 to 06.00 or 08.00 hours), under two different nutritional regimens with 100 or 50 g of protein/day. Furthermore, rates of degradation of rRNA and tRNA were calculated using values for these RNA catabolites. For comparison, the corresponding excretion of creatinine, which originates from the energy metabolism of muscle, and of 3-methylhistidine (m(3)His), which is an indicator of muscle protein degradation, was determined. Differences in excretion during the collection periods were tested using the Friedmann test. The excretion of psi, creatinine and m(3)His (micromol x h(-1) x kg(-1)) altered significantly (P <0.001) during the day. Medians were: for psi, 0.21 (morning), 0.19 (afternoon), 0.19 (evening) and 0.18 (night); for creatinine, 8.8, 8.4, 8.0 and 7.3 respectively; for m(3)His, 0.13, 0.11, 0.12 and 0.10 respectively. The excretion rates of m(2)(2)G and t(6)A (nmol x h(-1) x kg(-1)) altered, but not significantly, during the day; corresponding medians were: for m(2)(2)G, 9.0, 8.4, 8.0 and 8.4 respectively; for t(6)A, 4.3, 4.1, 3.9 and 3.9 respectively. From these results it can be concluded that, in order to assess the average daily rates of degradation of tRNA and rRNA using modified RNA catabolites, urine collection should be carried out quantitatively over 24 h periods. Likewise, the catabolites creatinine and m(3)His must be determined using 24 h urine samples when average daily excretion values are required.

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Chapter 2 Ribonucleosides in Biological Fluids by A High-Resolution Quantitative Rplc-UV Method

Cited by 10 publications

References 43 publications

Blood Plasma Pseudouridine in Patients with Malignant Proliferative Diseases

Blood Plasma Pseudouridine in Patients with Malignant Proliferative Diseases

Noninvasive Markers of Oxidative DNA Stress, RNA Degradation and Protein Degradation Are Differentially Correlated With Resting Metabolic Rate and Energy Intake in Children and Adolescents

Diurnal variation in the renal excretion of modified RNA catabolites in humans

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