Supramolecular spectral/visual detection of urinary polyamines through synergetic/competitive complexation with γ-CD and CB[7]

A supramolecular strategy for detecting the concentration of polyamines has been established through competitive/synergetic complexation among polyamines, CB[7], γ-CD, and pyrene derivatives, which allows for convenient, rapid, and high throughput spectral/visual detection of the concentration of urinary polyamines based on the switching on/off of the pyrene excimer fluorescence.

Cell kinetics and biochemical parameters in breast cancer

The study analyzes biochemical and cell kinetic parameters to characterize solid tumor growth in humans. The concentrations of polyamines, CEA, the thymidine labeling index (T.L.I.) and the mitotic index (M.I.) were determined on fragments of neoplastic tissue from 18 patients with breast carcinoma. Urinary polyamines were evaluated in the same patients. Two groups of patients were distinguished according to the median value of the T.L.I. In the group with high T.L.I., M.I. and tissue polyamines were significantly higher than in the group with low T.L.I., whereas tissue CEA was lower, though in a not statistically significant way. Urinary polyamines showed no variations between groups. These preliminary results showed that T.L.I. levels were higher in patients who relapsed during a 4-year follow-up than in patients achieving complete remission and remaining disease free. Results concerning polyamine concentration showed that the tissue polyamine level in breast carcinoma indicated proliferative activity, but this does not seem to be valuable for current prognostic purposes

Simple, sensitive assay of polyamines by high-performance liquid chromatography with electrochemical detection after post-column reaction with immobilized polyamine oxidase.

This simple, rapid liquid-chromatographic assay of urinary polyamines (putrescine, spermidine, spermine, and cadaverine) involves electrochemical detection with a post-column immobilized enzyme, polyamine oxidase (EC 1.4.3.6) from soybean seedlings. Polyamines are separated by isocratic ion-pairing reversed-phase chromatography, then enzymatically converted, with release of hydrogen peroxide, via the post-column reactor with immobilized polyamine oxidase; the hydrogen peroxide is detected by electrochemical oxidation on a platinum electrode. The detection limits for injected putrescine, spermidine, and spermine were 0.3, 0.5, 0.6, and 4 pmol, respectively, with linear ranges of two to three orders of magnitude. Reproducibility was also good, with CV values less than 7%. The efficiency of the immobilized enzyme column was not decreased after analysis of 300 urine samples. Putrescine and spermidine excretion in urine from patients with blood cancers and solid cancers was significantly increased.

Clinical usefulness of an enzymatic determination of total urinary polyamines, excluding cadaverine.

In one widely used enzymatic method for urinary polyamines, the total concentrations of four polyamines--putrescine, spermidine, spermine, and cadaverine--are determined. We report here a simple enzymatic method for measuring the total concentrations of urinary polyamines except cadaverine. The coefficients of variation (CV) for within-run measurements by this method were 4.3% (means = 17.2 mumol/L) and 1.5% (means = 66.5 mumol/L), between-run CVs were 4.8% (means = 16.8 mumol/L) and 1.8% (means = 67.5 mumol/L). The central 95% normal reference interval was 12.3-29.1 mumol/g creatinine for men and 14.1-36.8 mumol/g creatinine for women. In some cases, physiological variations in urinary polyamine excretion were large, mainly because of variations in cadaverine excretion, even in health. Pathological changes in polyamine production in the body may therefore be more easily shown by the excretion of total polyamines excluding cadaverine than by that including cadaverine.