Capillary electrophoresis in the evaluation of ischemic injury: Simultaneous determination of purine compounds and glutathione

Carlucci, Filippo; Tabucchi, Antonella; Biagioli, Bonizella; Sani, Guido; Lisi, Gianfranco; Maccherini, Massimo; Rosi, F.; Marinello, Enrico

doi:10.1002/(sici)1522-2683(20000501)21:8<1552::aid-elps1552>3.0.co;2-m

Cited by 34 publications

(12 citation statements)

References 33 publications

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“…Being the precursors of UA and the intermediates in purine nucleotide or deoxynuclotide metabolism, the determination of Xa and Hxa is important in biochemical and clinical diagnosis [18]. Various techniques have been developed to determine the purine derivatives, including enzymatic methods [19,20], HPLC [21][22][23][24], HPCE [25][26][27] and electrochemistry methods [28,29], among which enzymatic approaches are valuable analytical tools and offer sensitive and specific methods for the determination of Xa, Hxa and UA. However, the xanthine oxidase can oxidize both Hxa and Xa to UA making the specific determination difficult.…”

Section: Introductionsupporting

confidence: 86%

An inlaying ultra-thin carbon paste electrode modified with functional single-wall carbon nanotubes for simultaneous determination of three purine derivatives

Wang

Dong

2008

Sensors and Actuators B: Chemical

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Section: Introductionsupporting

confidence: 86%

An inlaying ultra-thin carbon paste electrode modified with functional single-wall carbon nanotubes for simultaneous determination of three purine derivatives

Wang

Dong

2008

Sensors and Actuators B: Chemical

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“…Nucleotides were evaluated as previously described [12]. The cells (5 × 10 6 ) were homogenized in 2.7 N perchloric acid in 0.5-ml tubes using a Sigma nylon motor pestle.…”

Section: Methodsmentioning

confidence: 99%

Critical Appraisal of the MTT Assay in the Presence of Rottlerin and Uncouplers

et al. 2009

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Rottlerin is a natural product isolated from Mallotus philippinensis. This polyphenolic compound, originally described as a selective inhibitor of PKCδ, can inhibit many other PKC-unrelated kinases and has a number of biological actions, including mitochondrial uncoupling effects. We recently found that Rottlerin inhibits the transcription factor nuclear factor κB in different cell types, causing downregulation of cyclin D1 and growth arrest. The present study was carried out to clarify the surprising lack of effect of Rottlerin on MCF-7 cell viability, assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) test. We found that Rottlerin causes overestimation of the MTT test, leading to inconsistent results between cell number and cell viability. Rottlerin, however, strongly differs from other antioxidant polyphenols, which directly reduce tetrazolium salts, since it does not exhibit any reactivity toward the tetrazolium salts in vitro nor does it modulate lactate dehydrogenase activity. The interference in the MTT assay occurred only in cultured cells, concomitantly with a decrease in the energy charge. Because the same MTT overestimation was observed in the presence of uncoupling agents, we conclude that the Rottlerin artifact is linked to its uncoupling action that, by accelerating oxidative chain, accidentally results in enhanced MTT reduction. These results suggest caution in the use of the MTT assay in the presence of Rottlerin and uncouplers in general.

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“…Aliquots were analysed by capillary electrophoresis (CE) for purine compounds (nucleotides, nucleosides and bases) and reduced/ oxidised glutathione, according to Carlucci et al [2] The same CE method was utilised for the determination of correlated enzyme activities: purine nucleoside phosphorylase (PNP), adenosine deaminase (ADA), and the two soluble isoforms of 5'-nucleotidase (e-Ns and c-N-II) AMP and IMP dependent respectively. Biopsies were obtained before explantation procedure (t1), after cold ischemic period (t2) and 30 min.…”

Section: Methodsmentioning

confidence: 99%

Liver Transplant: Adenosine Metabolism and Apoptosis

Carlucci

Marinello

Gerunda

et al. 2004

Nucleosides, Nucleotides and Nucleic Acids

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Apoptosis and necrosis coexist in ischemia-reperfusion (I/R) injury following organ transplant. During experimental liver transplant we evidenced a deep alteration in energy and antioxidant status. The activity of purine catabolic enzymes was also altered. Caspase-3 (C-3), protein tyrosine phosphatase (PTP) showed significative alterations that lead to DNA fragmentation. These findings could be of interest in new potential strategy to prevent and treat I/R injury.

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Capillary electrophoresis in the evaluation of ischemic injury: Simultaneous determination of purine compounds and glutathione

Cited by 34 publications

References 33 publications

An inlaying ultra-thin carbon paste electrode modified with functional single-wall carbon nanotubes for simultaneous determination of three purine derivatives

An inlaying ultra-thin carbon paste electrode modified with functional single-wall carbon nanotubes for simultaneous determination of three purine derivatives

Critical Appraisal of the MTT Assay in the Presence of Rottlerin and Uncouplers

Liver Transplant: Adenosine Metabolism and Apoptosis

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