José A. Pascual

Assessment of chronic exposure to cigarette smoke and its change during pregnancy by segmental analysis of maternal hair nicotine

This study aimed to investigate the association between biomarkers of fetal exposure to smoking during the whole pregnancy, nicotine in maternal and newborns hair samples, and quantitative measurement of smoking intake and exposure evaluated by maternal self-reported questionnaire. Study subjects were 150 mothers and their newborns from a hospital in Barcelona. A questionnaire including smoking habits was completed in the third trimester of pregnancy and on the day of delivery. Nicotine content was measured in two subsequent segments of maternal hair accounting for the first and last months of pregnancy, and in fetal hair. The geometric mean of nicotine concentration in maternal hair discriminated between nonexposition (3.84 and 2.80u2009ng/mg in distal and proximal hair segment, respectively) and exposition to cigarette smoke during pregnancy (6.06 and 4.30u2009ng/mg in distal and proximal hair segment, respectively) (P<0.05), and between these two classes and active smoking (14.40 and 11.08u2009ng/mg in distal and proximal hair segment, respectively). Maternal hair nicotine was able to differentiate levels of exposure to tobacco smoke and levels of intake. Nicotine concentration in hair from newborns did not differentiate between exposure and nonexposure to environmental tobacco smoke (ETS) in nonsmoking mothers. Finally, chronic exposure to cigarette smoke during pregnancy, assessed by maternal hair nicotine, correlated negatively with anthropometric parameters of newborns.

Neurotoxic Thioether Adducts of 3,4-Methylenedioxymethamphetamine Identified in Human Urine After Ecstasy Ingestion

3,4-Methylenedioxymethamphetamine (MDMA, Ecstasy) is a widely misused synthetic amphetamine derivative and a serotonergic neurotoxicant in animal models and possibly humans. The underlying mechanism of neurotoxicity involves the formation of reactive oxygen species although their source remains unclear. It has been postulated that MDMA-induced neurotoxicity is mediated via the formation of bioreactive metabolites. In particular, the primary catechol metabolites, 3,4-dihydroxymethamphetamine (HHMA) and 3,4-dihydroxyamphetamine (HHA), subsequently cause the formation of glutathione and N-acetylcysteine conjugates, which retain the ability to redox cycle and are serotonergic neurotoxicants in rats. Although the presence of such metabolites has been recently demonstrated in rat brain microdialysate, their formation in humans has not been reported. The present study describes the detection of 5-(N-acetylcystein-S-yl)-3,4-dihydroxymethamphetamine (N-Ac-5-Cys-HHMA) and 5-(N-acetylcystein-S-yl)-3,4-dihydroxyamphetamine (N-Ac-5-Cys-HHA) in human urine of 15 recreational users of MDMA (1.5 mg/kg) in a controlled setting. The results reveal that in the first 4 h after MDMA ingestion ∼0.002% of the administered dose was recovered as thioether adducts. Genetic polymorphisms in CYP2D6 and catechol-O-methyltransferase expression, the combination of which are major determinants of steady-state levels of HHMA and 4-hydroxy-3-methoxyamphetamine, probably explain the interindividual variability seen in the recovery of N-Ac-5-Cys-HHMA and N-Ac-5-Cys-HHA. In summary, the formation of neurotoxic thioether adducts of MDMA has been demonstrated for the first time in humans. The findings lend weight to the hypothesis that the bioactivation of MDMA to neurotoxic metabolites is a relevant pathway to neurotoxicity in humans.

Evaluation of immunoassays for the measurement of insulin-like growth factor-I and procollagen type III peptide, indirect biomarkers of recombinant human growth hormone misuse in sport

Abstract Insulin-like growth factor-I (IGF-I) and procollagen type III peptide (P-III-P) have been proposed as indirect biomarkers for the detection of the misuse of recombinant human growth hormone in sport. An extended intra- and inter-laboratory validation of commercially available immunoassays was carried out. For total IGF-I, two radioimmunoassay (RIA) kits (IGF-I/RIA1, Nichols Institute Diagnostics and IGF-I/RIA2, Mediagnost) and one enzyme-linked immunosorbent assay (ELISA) (R&D) were evaluated. For P-III-P, two RIA kits (P-III-P/RIA3, Cis-bioInternational and P-III-P/RIA4, Orion Diagnostica) were studied. The intra-laboratory precision and accuracy values for all IGF-I assays were better than 15%. The IGF-I/ELISA showed the lowest limit of quantification (LOQ) and its calibration curve covered the range of concentrations found in human serum samples. Higher agreement between laboratory results was obtained for IGF-I/ELISA and IGF-I/RIA1. Low inter-technique correlation was obtained for the three assays; the only comparable results were obtained between IGF-I/ELISA and IGF-I/RIA1. For P-III-P, intra-laboratory precision and accuracy values better than 15% were obtained for both assays in almost all cases. The calibration curve for P-III-P/RIA4 covered the range of concentrations of serum samples, while 30% of the values for P-III-P/RIA3 were below the calibration sample with the lowest concentration. Inter-laboratory correlation was also higher for P-III-P/RIA4. In summary, ELISA and RIA4 were the most suitable assays for measurement of IGF-I and P-III-P, respectively, in serum samples. However, the validation studies carried out show the need for harmonization of immunoassay parameters to improve the reproducibility and comparability of results between different laboratories and in different studies.

Recombinant erythropoietin found in seized blood bags from sportsmen

During an anti-doping investigation, the Spanish Guardia Civil confiscated blood bags from elite sportsmen. A novel immuno-purification method demonstrated that plasma samples with elevated erythropoietin (EPO) contained recombinant material (rEPO). This shows that rEPO is used before autologous blood transfusions and that rEPO analysis in plasma can be reliably addressed.

Intermittent hypoxia exposure in a hypobaric chamber and erythropoietin abuse interpretation

Abstract The aim of this study was to assess the effect of intermittent hypoxia exposure on direct and indirect methods used to evaluate recombinant human erythropoietin (rhEPO) misuse. Sixteen male triathletes were randomly assigned to either the intermittent hypoxia exposure group (experimental group) or the control normoxic group (control group). The members of the experimental group were exposed to simulated altitude (from 4000 to 5500 m) in a hypobaric chamber for 3 h per day, 5 days a week, for 4 weeks. Blood and urine samples were collected before and after the first and the final exposures, and again 2 weeks after the final exposure. While serum EPO significantly increased after the first [from a mean 8.3 IU · l−1 (s = 3.2) to 16.6 IU · l−1 (s = 4.7)] and final exposures [from 4.6 IU · l−1 (s = 1.4) to 24.8 IU · l−1 (s = 9.3)], haemoglobin, percentage of reticulocytes, and soluble transferrin receptor were not elevated. Second-generation ON/OFF models (indirect rhEPO misuse detection) were insensitive to intermittent hypoxia exposure. The distribution of the urinary EPO isoelectric profiles (direct rhEPO misuse detection) was altered after intermittent hypoxia exposure with a slight shift towards more basic isoforms. However, those shifts never resulted in misinterpretation of results. The intermittent hypoxia exposure protocol studied did not produce any false-positive result for indirect or direct detection of rhEPO misuse in spite of the changes in EPO serum concentrations and urinary EPO isoelectric profiles, respectively.

Research Spotlight: Bioanalysis and Analytical Services Research Group at The Municipal Institute for Medical Research IMIM-Hospital del Mar, Spain

Analytical laboratories involved in health-related research are becoming a fundamental part of the advancement of science in this field. Of particular interest to clinical, legal, toxicological, forensic and environmental matters is the analysis of drugs and medications present in biological fluids of consumers or exposed subjects. The established sensitive and reliable work of sports drug-testing laboratories represents an interesting example of a multidisciplinarity approach toward widespread bioanalytical problems. The experiences reported in this article will be of general interest, especially for analysts studying the determination of substances in biological material.