The precise radioimmunoassay of adenosine: Minimization of sample collection artifacts and immunocrossreactivity

Linden, Joel; Taylor, Heidi E.; Feldman, Marc D.; Woodward, E. Baylor; Ayers, Carlos R.; Ripley, Marcia L.; Iflah, Shulah; Patel, Amrat

doi:10.1016/0003-2697(92)90335-5

Cited by 15 publications

(10 citation statements)

References 20 publications

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“…Plasma was collected from RR MS patients (n ϭ 6) and healthy control subjects (n ϭ 6) according to established procedures, 28 and adenosine levels were determined as previously described. 29 Briefly, whole blood was drawn and concurrently mixed at approximately a 1:1 ratio with an ice-cold adenosine stop solution containing 10 mM of erythro-9-(2hydroxy-3-nonyl)adenine hydrochloride, 10 mM of ethylenediaminetetraacetic acid and EGTA, 10 mM of d-L-␣glycerophosphate, 0.2 mM of dipyridamole, 0.1% heparin, and 150 mM of NaCl.…”

Section: Plasma Adenosine Levelsmentioning

confidence: 99%

Dysregulation of adenosine A1 receptor-mediated cytokine expression in peripheral blood mononuclear cells from multiple sclerosis patients

et al. 1999

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Cytokines, including tumor necrosis factor‐α (TNFα) and interleukin‐6 (IL‐6), have been implicated in the pathogenesis of multiple sclerosis (MS). The production and release of these cytokines are regulated in part by specific purinergic (adenosine) cell surface receptors. To determine the extent to which the adenosine A1 receptor influenced cytokine expression in peripheral blood mononuclear cells (PBMCs) from MS and control patients, we measured plasma adenosine and TNFα levels, A1 receptor messenger RNA (mRNA) and protein amounts, and the effects of activation of A1 receptors on TNFα and IL‐6 production by PBMCs. Plasma levels of TNFα were significantly higher and adenosine levels were significantly lower in MS patients compared with control subjects. Levels of TNFα and IL‐6 in mitogen‐stimulated PBMC culture supernatants from MS patients or control patients were similar. Conversely, treatment of PBMCs with the adenosine A1 receptor agonist R‐phenylisopropyladenosine (R‐PIA) (1 μM) significantly inhibited mitogen‐stimulated production of TNFα but not IL‐6 in control subjects and significantly inhibited production of IL‐6 but not TNFα in MS patients. The effects of R‐PIA were selectively blocked by the A1 receptor antagonist 8‐cyclopentyl‐1,3‐dipropylxanthine (DPCPX). A1 receptor protein levels were decreased significantly in PBMCs from MS patients. Taken together, these results suggest that decreased levels of adenosine and its A1 receptor modulate TNFα and IL‐6 levels and may contribute to the pathogenesis of MS. Ann Neurol 1999;45:633–639

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Section: Plasma Adenosine Levelsmentioning

confidence: 99%

Dysregulation of adenosine A1 receptor-mediated cytokine expression in peripheral blood mononuclear cells from multiple sclerosis patients

et al. 1999

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“…A radioimmunoassay (mean of 4 assays) of urinary adenosine was used. 21 Plasma B 12 and blood folate levels were measured for each sample using a radioisotope competition assay (Ciba Corning, East Walpole, Mass) and a methotrexate-resistant Lactobacillus casei assay, respectively. 22 Urinary creatinine levels were measured in duplicate using the Sigma Diagnostics Creatinine Kit (Procedure 555; Sigma Chemical Co, St Louis, Mo).…”

Section: Laboratory Methodsmentioning

confidence: 99%

Urinary Adenosine and Aminoimidazolecarboxamide Excretion in Methotrexate-Treated Patients With Psoriasis

Baggott¹,

Morgan²,

Sams³

et al. 1999

Arch Dermatol

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“…It is to be noted that although immunology techniques appear to be more sensitive 21,22 only HPLC enables both adenosine and its derivatives to be measured in the same sample. Quantities determined with the two HPLC methods showed that the correlation between the two techniques was good.…”

Section: Resultsmentioning

confidence: 99%

“…13 5'nucleotidase is inhibited by (α,β methylene adenosine 5' diphosphate (AOPCP). 19 Once the already delicate sampling phase has terminated, several quantitative assay methods for adenosine and its metabolites have been described, including fluorimetric assay, 20 radioimmunoassay 21,22 or HPLC. 15,16 The sampling methods using the ultra-rapid mixing of blood and stopping solution are composed of a double syringe 17 or a double lumen catheter.…”

Section: Introductionmentioning

confidence: 99%

“…14 Finally, EHNA (erythro-9(2-hydroxy-3 nonyl)adenine and 2'deoxycorformycin are potent inhibitors of adenosine deaminase. [23][24][25][26][27][28][29] These techniques are long and tedious since they require either extraction followed by radioimmunoanalysis 22 freeze-drying cycles before HPLC identification and quantitation. 18 We have recently described a sampling technique using a vacuum tube for the simultaneous aspiration of blood and the stopping solution, and their rapid mixing.…”

Section: Introductionmentioning

confidence: 99%

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Development of an HPLC/Diode Array Detector Method for the Determination of Human Plasma Adenosine Concentrations

Guieu¹,

Sampiéri²,

Bechis³

et al. 1999

Journal of Liquid Chromatography & Related Technologies

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Assays of adenosine and its derivatives in biological fluids involve fluorimetric, radioimmunological or chromatographic analyses. Techniques currently used are tedious because they involve either an extraction using immunological methods or one or two freeze drying cycles when using high performance liquid chromatography (HPLC). In this context, we describe a "quick" HPLC method using a diode array detector for the spectral analysis and quantitation of adenosine and its derivatives in less than two hours following sampling. We compared our results to those obtained with an HPLC technique coupled to UV detection. Downloaded by [UQ Library] at 02:36 01 November 2014 ORDER REPRINTS 1830 GUIEU ET AL.Although there is a good correlation between the two techniques, the "quick" method provide values that are on average 30% higher than those obtained with the classical method. The absence of a lyophilization step in the classical method could explain this difference. The utilization of an HPLC system coupled to a diode array detector leads to a rapid and reliable assay of adenosine and its derivatives. The possibility of having a rapid assay method available could be very useful in cases of certain cardiovascular or neurological disorders.

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The precise radioimmunoassay of adenosine: Minimization of sample collection artifacts and immunocrossreactivity

Cited by 15 publications

References 20 publications

Dysregulation of adenosine A1 receptor-mediated cytokine expression in peripheral blood mononuclear cells from multiple sclerosis patients

Dysregulation of adenosine A1 receptor-mediated cytokine expression in peripheral blood mononuclear cells from multiple sclerosis patients

Urinary Adenosine and Aminoimidazolecarboxamide Excretion in Methotrexate-Treated Patients With Psoriasis

Development of an HPLC/Diode Array Detector Method for the Determination of Human Plasma Adenosine Concentrations

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