Are Carbohydrate-Deficient Transferrin Assays Useful for the Detection of Recurrent 'Binge Drinking' in Children with an Alcohol Intoxication in the Emergency Department?

AIMS: The aim of this study was to evaluate different carbohydrate-deficient transferrin (CDT) assays for the detection of recurrent excessive alcohol abuse in adolescents prior to acute alcohol intoxication. METHODS: Data on drinking behaviour and CDT levels of adolescents (13-18 years) registered at the outpatient clinic for youth and alcohol at three major district general hospitals in the Netherlands were retrospectively collected. CDT and disialotransferrin (DST) levels of binge-drinking teenagers were compared with non-binge-drinking teenagers. RESULTS: In total 198 samples were collected for the N Latex CDT method (N = 83), no differences were found in mean CDT levels for binge versus non-binge drinkers (P = 0.8). The Helander HPLC (N = 78) showed significantly higher values for binge drinkers than for non-binge drinkers (mean 1.20%DST, SD 0.28 and mean 1.01%DST, SD 0.31, respectively (P = 0.01)). The Recipe ClinRep method (N = 37) also showed significantly higher values for binge drinkers (mean 1.17%DST, SD 0.36 and mean 0.89%DST, SD 0.34, respectively (P = 0.03)). CONCLUSION: With the Helander HPLC method and the Recipe ClinRep assay higher levels are measured in binge drinkers than in non-binge drinkers.

Quantification of urinary o,o'-dityrosine, a biomarker for oxidative damage to proteins, by high performance liquid chromatography with triple quadrupole tandem mass spectrometry. A comparison with ion-trap tandem mass spectrometry.

We recently described an isotope dilution reversed-phase liquid chromatography-atmospheric pressure chemical ionization-ion-trap-tandem mass spectrometry (HPLC-APCI-MS/MS) method for the quantitative determination of oxidized amino acids in human urine, including o,o'-dityrosine, a specific marker of protein oxidation. In the present study, we investigated the possibility to use a triple quadrupole instrument for the analysis of this biomarker in urine. The two instruments were compared in terms of sensitivity, specificity and reproducibility. Results showed that the triple quadrupole instrument reaches 2.5-fold higher sensitivity (LOD=0.01 microM) compared to the previously used ion-trap instrument. Precision of the present assay is as follows: in-day variation is 4.6% and inter-day variation is 17%. The currently developed method was applied to a group of smoker urine samples. The mean urinary o,o'-dityrosine concentration was 0.08+/-0.01 microM. Expressed per urinary creatinine concentration, this corresponds to 10.1+/-0.4 micromol/mol creatinine. This is comparable to the previously reported values of 5.8+/-0.3 micromol/mol creatinine in non-smokers night-time urines, and 12.3+/-5 micromol/mol creatinine in day-time urines measured by the ion-trap instrument.

Spread supplemented with moderate doses of vitamin E and carotenoids reduces lipid peroxidation in healthy, nonsmoking adults.

BACKGROUND: High doses of vitamin E have been shown to decrease lipid peroxidation in persons under oxidative stress. At present, the data are insufficient to predict whether lower doses offer the same benefit in healthy persons. OBJECTIVE: We studied the effect of moderate doses of a combination of vitamin E and carotenoids, incorporated into a food product, on markers of antioxidant status and lipid peroxidation in healthy persons. DESIGN: One hundred five healthy adults were randomly, evenly assigned in this double-blind, placebo-controlled, parallel, 11-wk intervention study. After a 2-wk stabilization period during which the subjects consumed a commercial unfortified spread, the subjects consumed 25 g/d of spread containing 43 mg alpha-tocopherol equivalents (alpha-TE; 2-3 fold the US dietary reference intake) and 0.45 mg carotenoids (spread A), 111 mg alpha-TE and 1.24 mg carotenoids (spread B), or 1.3 mg RRR-alpha-tocopherol without carotenoids (spread C). RESULTS: In subjects consuming spread A, plasma alpha-tocopherol concentrations increased 31% to 32 micromol/L, with small but significant increases in concentrations of alpha-carotene and lutein. This resulted in LDL with significantly higher total antioxidant capacity (17%) and an increased resistance to oxidation, as determined by lag time (18%). These improvements were dose dependent: larger increases in these variables were observed in subjects consuming spread B. Furthermore, consumption of spread B significantly reduced concentrations of the plasma lipid peroxidation biomarker F(2 alpha)-isoprostane (15%). CONCLUSION: The consumption of food products containing moderate amounts of vitamin E and carotenoids can lead to measurable and significant improvements in antioxidant status and biomarkers of oxidative stress in healthy persons.

Assessment of the efficacy of functional food ingredients-introducing the concept "kinetics of biomarkers".

Functional foods are "foods and beverages with claimed health benefits based on scientific evidence". Health claims need to be substantiated scientifically. The future of functional foods will heavily rely on proven efficacy in well-controlled intervention studies with human volunteers. In order to have the maximum output of human trials, improvements are needed with respect to study design and optimization of study protocols. Efficacy at realistic intake levels needs to be established in studies with humans via the use of suitable biomarkers, unless the endpoint can be measured directly. The human body is able to deal with chemical entities irrespective of their origin, and the pharmaceutical terms "absorption, distribution, metabolism and excretion" have their equivalent when biomarkers are concerned. Whereas only "diurnal variation" or "circadian rhythm" is sometimes considered, little attention is paid to "kinetics of biomarkers". "Kinetics of biomarkers" comprises "formation, distribution, metabolism and excretion". However, this is at present neither an established science nor common practice in nutrition research on functional foods. As a consequence, sampling times and matrices, for example, are chosen on the basis of historical practice and convenience (for volunteers and scientists) but not on the basis of in depth insight. The concept of kinetics of biomarkers is illustrated by a variety of readily comprehensible examples, such as malaria, cholesterol, polyphenols, glutathione-S-transferase alpha, F2-isoprostanes, interleukin-6, and plasma triacylglycerides.

A new method for measuring oxidative stress in claudicants during strenuous exercise using free radical derivatives of antipyrine as indicators: a pilot study.

Patients with intermittent claudication disease suffer from temporary lack of oxygen in the legs, caused by narrowing of arteries, resulting in ischemia and followed by reperfusion. The degree of oxidative stress present in 16 patients during strenuous exercise was determined using several indicators. Two derivatives of an exogenous marker, antipyrine (AP), (ie, p-hydroxyantipyrine, p-APOH, and o-hydroxyantipyrine, o-APOH), were assayed in plasma using HPLC-tandem-MS. Plasma malondialdehyde (assayed as thiobarbituric acid reactive species, TBARS) was also determined. The branchial/ankle blood pressure index (b-a index) was used to assess the severity of intermittent claudication disease, and plasma lactate concentration was also measured as an indicator of the ischemic situation. Plasma TBARS level did not change significantly after exercise. During the ischemic situation as well as during reperfusion, both free radical derivatives of antipyrine increased significantly in plasma (p < 0.01). Because p-APOH is also formed enzymatically in humans, the plasma ratio of o-APOH to AP appeared to be the most specific marker for oxidative stress in patients with intermittent claudication.

Measurement of ischaemia-reperfusion in patients with intermittent claudication using NMR-based metabonomics.

Intermittent claudication has proved to be a good in vivo model for ischaemia-reperfusion. For assessment of ischaemia-reperfusion damage, the known biochemical markers all have disadvantages with respect to sensitivity and interference with other physiological events. In this work, we studied the metabolic effects of ischaemia-reperfusion in patients with intermittent claudication, and the effects of vitamin C and E intervention, using both traditional biochemical measurements and 1H-NMR-based metabonomics on urine and plasma. The 1H-NMR spectra were subjected to multivariate modelling using principal components discriminant analysis, and the observed clusters were validated using joint deployment of univariate analysis of variance and Tukey-Kramer honestly significant difference (HSD) testing. The study involved 14 patients with intermittent claudication and three healthy volunteers, who were monitored during a walking test, before and after a vitamin C/E intervention, and after a washout period. The effect of exercise was only observable for a limited number of biochemical markers, whereas 1H NMR revealed an effect in line with anaerobic ATP production via glycolysis in exercising (ischaemic) muscle of the claudicants. Thus, the beneficial effect of vitamins C and E in claudicants was more pronounced when observed by metabonomics than by traditional biochemical markers. The main effect was more rapid recovery from exercise to resting state metabolism. Furthermore, after intervention, claudicants tended to have lower concentrations of lactate and glucose and several other citric acid cycle metabolites, whereas acetoacetate was increased. The observed metabolic changes in the plasma suggest that intake of vitamin C/E leads to increased muscle oxidative metabolism.

Olive oils high in phenolic compounds modulate oxidative/antioxidative status in men.

The aim of the present study was to evaluate whether olive oils high in phenolic compounds influence the oxidative/antioxidative status in humans. Healthy men (n = 12) participated in a double-blind, randomized, crossover study in which 3 olive oils with low (LPC), moderate (MPC), and high (HPC) phenolic content were given as raw doses (25 mL/d) for 4 consecutive days preceded by 10-d washout periods. Volunteers followed a strict very low-antioxidant diet the 3 d before and during the intervention periods. Short-term consumption of olive oils decreased plasma oxidized LDL (oxLDL), 8-oxo-dG in mitochondrial DNA and urine, malondialdehyde in urine (P < 0.05 for linear trend), and increased HDL cholesterol and glutathione peroxidase activity (P < 0.05 for linear trend), in a dose-dependent manner with the phenolic content of the olive oil administered. At d 4, oxLDL after MPC and HPC, and 8-oxo-dG after HPC administration (25 mL, respectively), were reduced when the men were in the postprandial state (P < 0.05). Phenolic compounds in plasma increased dose dependently during this stage with the phenolic content of the olive oils at 1, 2, 4, and 6 h, respectively (P < 0.01). Their concentrations increased in plasma and urine samples in a dose-dependent manner after short-term consumption of the olive oils (P < 0.01). In conclusion, the olive oil phenolic content modulated the oxidative/antioxidative status of healthy men who consumed a very low-antioxidant diet.