Hypoxanthine-guanine phosphoribosyltransferase deficiency and erythrocyte synthesis of pyridine coenzymes

Micheli, Vanna; S, Sestini; Rocchigiani, Marina; Jacomelli, Gabriella; Manzoni, Francesca; Peruzzi, Luana; Gathof, Birgit; Zammarchi, Enrico; Pompucci, G

doi:10.1016/s0024-3205(99)00205-2

Cited by 31 publications

(16 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The described HPLC method, initially devised to measure purines, pyrimidines and other metabolites in plasma and urines (Micheli et al 1999), allows the identification and quantification of several compounds, including creatinine, uric acid, tyrosine, hypoxanthine, xanthine, HGA, phenylalanine and tryptophan ( Fig.2a and b).…”

Section: Hplc Analysis: Methods Validationmentioning

confidence: 99%

See 1 more Smart Citation

Quick Diagnosis of Alkaptonuria by Homogentisic Acid Determination in Urine Paper Spots

et al. 2016

View full text Add to dashboard Cite

Objectives: Two methods are described for homogentisic acid (HGA) determination in dried urine spots (DUS) on paper from Alkaptonuria (AKU) patients, devised for quick early diagnosis. AKU is a rare autosomal recessive disorder caused by deficiency of homogentisate 1,2-dioxygenase, yielding in accumulation of HGA. Its massive excretion causes urine darkening by exposure to air or alkalinization, and is a diagnostic marker. The deposition of polymers produced after HGA oxidation within the connective tissues causes ochronotic arthritis, a degenerative joint disease manifesting in adulthood and only rarely in childhood. No early diagnosis is usually accomplished, awareness following symptom development.Design and methods: Two methods were designed for HGA determination in DUS: (1) a rapid semi-quantitative reliable method based on colour development in alkali and quantification by comparison with dried paper spots from HGA solutions of known concentration and (2) a quantitative and sensitive HPLC-linked method, previously devised for purine and pyrimidine analysis in urine and plasma.Results: Colour intensity developed by DUS after alkali addition was proportional to HGA concentration, and calculated amounts were in good agreement with quantitative analysis performed by RP-HPLC on DUS and on urines as such.Conclusions: DUS, often used for different diagnostic purpose, are easily prepared and safely delivered. The simple and quick colour method proposed provides reliable HGA assessment and is fit for large screening. HGA concentration determined in 10 AKU patient DUS by both methods 1 and 2 was in agreement with direct urine assay and in the range reported by literature.A reliable HGA quantification based on colour development in paper urine spots is validated by HPLC-linked HGA quantification, and proposed as a quick diagnostic tool for AKU patients.

show abstract

Section: Hplc Analysis: Methods Validationmentioning

confidence: 99%

“…A previously described HPLC method, initially devised to detect purines, pyrimidines, and other metabolites in plasma and urines (Micheli et al 1999), was used to measure HGA in DUS, in HGA solution spots and in urines.…”

Section: Hplc Analysismentioning

confidence: 99%

Quick Diagnosis of Alkaptonuria by Homogentisic Acid Determination in Urine Paper Spots

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Acid supernatant was brought to neutrality with 3.5 M K 2 CO 3 and immediately processed by HPLC or stored at -20°C until used. HPLC elution was conducted as previously described for erythrocyte nucleotide analysis with minor modifications [19].…”

Section: Analysis Of Purine and Pyrimidine Compoundsmentioning

confidence: 99%

Identification of the Nucleotidase Responsible for the AMP Hydrolysing Hyperactivity Associated with Neurological and Developmental Disorders

et al. 2007

View full text Add to dashboard Cite

Nucleoside monophosphate phosphohydrolases comprise a family of enzymes dephosphorylating nucleotides both in intracellular and extracellular compartments. Members of this family exhibit different sequence, location, substrate specificity and regulation. Besides the ectosolic 5'-nucleotidase, several cytosolic and one mitochondrial enzymes have been described. Nevertheless, researchers refer any AMP-dephosphorylating activity to as 5'-nucleotidase, lacking a more accurate identification. Increase of AMP hydrolysing activity has been associated with neurological and developmental disorders. The identification of the specific enzyme involved in these pathologies would be fundamental for the comprehension of the linkage between the enzyme activity alteration and brain functions. We demonstrate that the described neurological symptoms are associated with increased ectosolic 5'-nucleotidase activity on the basis of radiochemical assays and immunoblotting analysis. Furthermore, present data evidence that the assay conditions normally applied for the determination of cytosolic 5'-nucleotidases activity in crude extracts are affected by the presence of solubilised ectosolic nucleotidase.

show abstract

“…An Ultrasphere C-18 (Beckman) column (4.6 × 25 mm, 5 m particle size) was used. The elution was performed routinely according to Stocchi et al [18], and, occasionally, to support the identification of chromatographic peaks, according to Micheli et al [19]. To test the toxicity of the added compounds, which, as previously reported [20], depends on initial cell density, a parallel experiment was performed in which labeled dAdo was substituted by the unlabeled compound.…”

Section: Hplc Analysismentioning

confidence: 99%

“…Another radioactive peak (b) was visible, the identification of which is not certain. In an attempt to identify the compound, the same sample was analyzed using a different elution system [19]. The compound does not correspond to dIno, but it is likely a nucleotide derivative, even though a reliable identification was not achieved.…”

Section: Hplc Analysismentioning

confidence: 99%

2′‐Deoxyadenosine causes apoptotic cell death in a human colon carcinoma cell line

Giannecchini

D'Innocenzo

Pesi

et al. 2003

J Biochem & Molecular Tox

View full text Add to dashboard Cite

The combination of 2'-deoxyadenosine and 2'-deoxycoformycin is toxic for the human colon carcinoma cell line LoVo. In this study we investigated the mode of action of the two compounds and have found that they promote apoptosis. The examination by fluorescence microscopy of the cells treated with the combination revealed the characteristic morphology associated with apoptosis, such as chromatin condensation and nuclear fragmentation. The occurrence of apoptosis was also confirmed by the release of cytochrome c and the proteolytic processing of procaspase-3 in cells subjected to the treatment. To exert its triggering action on the apoptotic process, 2'-deoxyadenosine enters the cells through an equilibrative nitrobenzyl-thioinosine-insensitive carrier, and must be phosphorylated by intracellular kinases. Indeed, in the present work we demonstrate by analysis of the intracellular metabolic derivatives of 2'-deoxyadenosine that, as suggested by our previous findings, in the incubation performed with 2'-deoxyadenosine and 2'-deoxycoformycin, an appreciable amount of dATP was formed. Conversely, when also an inhibitor of adenosine kinase was added to the incubation mixture, dATP was not formed, and the toxic and apoptotic effect of the combination was completely reverted.

show abstract

Hypoxanthine-guanine phosphoribosyltransferase deficiency and erythrocyte synthesis of pyridine coenzymes

Cited by 31 publications

References 28 publications

Quick Diagnosis of Alkaptonuria by Homogentisic Acid Determination in Urine Paper Spots

Quick Diagnosis of Alkaptonuria by Homogentisic Acid Determination in Urine Paper Spots

Identification of the Nucleotidase Responsible for the AMP Hydrolysing Hyperactivity Associated with Neurological and Developmental Disorders

2′‐Deoxyadenosine causes apoptotic cell death in a human colon carcinoma cell line

Contact Info

Product

Resources

About