MDMA (ecstasy) delays pubertal development and alters sperm quality after developmental exposure in the rat.

3,4-Methylenedioxymethamphetamine, MDMA or "ecstasy" is consumed mainly by young population at childbearing age. Therefore, there may be a risk of exposure of some pregnant women. The effects of the developmental exposure to MDMA on the sexual development and long-term sexual behaviour/fertility were assessed in Sprague-Dawley rats. MDMA was administered subcutaneously at 0 (control), 0.5, 5 and 10 mg/kg to female rats once a day, three consecutive days a week during 10 weeks, including gestation and lactation. The male offspring was evaluated for sexual maturation and mated with untreated sexually receptive females to evaluate the mating and pregnancy rates. Hormonal, haematological, biochemical, histological, genotoxicological and testicular and sperm parameters were also evaluated. A significant higher incidence of DNA damage in sperm and interstitial oedema in testes was found. There was also a significant and dose-related decrease in sperm count and a significant decrease in sperm motility at all doses. A significant delay in preputial separation onset in all treated groups was observed. This study reports by the first time an alteration of spermatogenesis after in utero and lactation MDMA exposure in the rat.

La coordinación entre profesores de fisiología y toxicología: un caso práctico en la Facultad de Farmacia de la Universidad de Barcelona / Physiology and toxicology teachers’ coordination. A case study in the Faculty of Pharmacy, University of Barcelona

El aprendizaje es un proceso continuo que no debería finalizar una vez aprobada una determinada asignatura. En cualquier estudio universitario hay muchas materias que, para su comprensión, requieren de conocimientos adquiridos previamente en otras. En los estudios de Farmacia del plan 2002, los profesores de toxicología habían constatado que los estudiantes de dicha asignatura no recordaban conceptos básicos cursados en asignaturas de semestres anteriores. La asignatura de toxicología necesita para su comprensión conocimientos de, entre otras materias, fisiología y fisiopatología. Por esta razón se planteó la necesidad de hacer una actuación conjunta entre los profesores de Fisiología y Toxicología.Los objetivos de este proyecto fueron: a) Identificación de los contenidos fisiológicos y fisiopatológicos que los alumnos deben conocer para el seguimiento de la asignatura de toxicología. b) Unificación terminológica. c) Realización de un conjunto de preguntas básicas sobre estos contenidos. d) Detección, a través de estas preguntas, de los temas o grupos de temas con porcentajes más altos de respuestas incorrectas. e) Detectar los temas de fisiología y fisiopatología en los que hay que hacer más hincapié para favorecer el seguimiento de toxicología. En esta comunicación se describe la experiencia y los resultados obtenidos(AU)

Learning is a continuous process that should be still performed once a particular subject has been passed. In the university, many subjects require prior knowledge of others subjects for better understanding. During the Pharmacy curriculum of 2002, toxicology teachers observed that students did not seem to remember the basic concepts presumably acquired in previous semesters. For example, for the toxicology subject, students should have basic knowledge of physiology and pathophysiology. For this reason, a joint action among physiology and toxicology teachers was considered.The objectives of this project were: a) Identifying the physiological and pathophysiological aspects that students should know to follow the toxicology course. b) Reaching agreement over the common terminology. c) Executing a set of basic questions about these physiological and pathophysiological aspects. d) Detecting through these questions, the topics with a highest percentage of incorrect answers. e) Identifying which physiology and pathophysiology topics should be emphasized to encourage students to follow the toxicology subject. This communication describes the experience and outcomes of this project(AU)

Biological monitoring of environmental exposure to manganese in blood samples from residents of the city of Barcelona, Spain.

Serum manganese levels were determined in 250 healthy subjects (122 men and 128 women) living in Barcelona in northeastern Spain. The study was designed to assess the reference levels for serum manganese and to investigate its relationship to age and sex. The age distribution ranged from 15 to 90 years. The assays were performed by means of a graphite furnace atomic absorption spectrometry. The geometric mean of serum manganese concentration was 1.1 microg/l, ranging from 0.3 microg/l to 2.5 microg/l. In almost every case, the 95th upper percentiles of this element were < 1.8 microg/l. No correlation between the concentration of manganese and sex could be established, but in the younger population the manganese levels were nearly three times higher than the results obtained in the older population.

Optimized procedure for lamotrigine analysis in serum by high-performance liquid chromatography without interferences from other frequently coadministered anticonvulsants.

The authors have developed a simple isocratic high-pressure liquid chromatographic (HPLC) assay for the simultaneous determination of lamotrigine and other frequently coadministered antiepileptic drugs in serum samples. Lamotrigine extraction was performed on a reversed-phase Oasis HBL preparation column. The eluates containing butalbital as internal standard were separated with a 7-microm Chromsystems C18 250 x 4.0 mm I.D. reversed-phase column at a temperature of 40 degrees C using a mobile phase consisting of pH 3.8 phosphate-acetonitrile buffer (55:45, v/v), at a flow rate of 0.8 mL/min. Ultraviolet detection was carried out at 210 nm. Measurement of the peak:height ratio allowed quantitative determination of the samples. The method was linear over a concentration range of 0.2 to 20 microg/mL for lamotrigine. Recovery was >90%. Within-day and between-day coefficients of variation ranged from 1.8% to 6.7%. The mean lamotrigine concentration was 8.01 +/- 5.63 microg/mL. After studying sera from 130 patients treated with lamotrigine the authors confirmed that associated antiepileptic therapy affected the serum lamotrigine levels, which were significantly higher in patients under valproic acid treatment.

Blood chromium determination in assessing reference values in an unexposed Mediterranean population.

Plasma chromium levels were determined in 243 healthy subjects. The study group consisted of 134 men and 109 women, ages 19-71 yr, all residing in Barcelona in northeastern Spain. The study was designed to assess the reference levels for plasma chromium and to investigate its relationships to age and sex. The assays were performed by means of a graphite-furnace atomic absorption spectrometer. The mean plasma chromium concentration was 3.01 +/- 1.45 nmol/L, ranging from 0.6 to 6 nmol/L. The upper reference values in the 0.95 percentile for this population was 5 nmol/L. No significant differences were observed with respect to the subjects' sex.

Serum and urine fluoride concentration: relationships to age, sex and renal function in a non-fluoridated population.

Serum and urine fluoride levels were determined in 250 healthy subjects (15-90 years, 122 men and 128 women) residing in Catalonia, Spain, and in 150 patients (20-81 years, 84 men and 66 women) with chronic renal failure undergoing regular dialysis treatment, living in the same geographical area, to determine normal range and to investigate its relationships to age, sex and renal function. Serum and urine fluoride were determined by a fluoride ion specific electrode system. Mean (+/- S.D.) serum fluoride concentration was 17.5 +/- 9.5 micrograms/l, ranging from 1 to 47 micrograms/l, in the control group and 58 +/- 31 micrograms/l, ranging from 28 to 185 micrograms/l, in renal patients. Urine fluoride concentration in the healthy group was 671 +/- 373 micrograms/24 h, ranging from 156 to 1900 micrograms/24 h. Fluoride status in the patient group was significantly greater than the control group. There was significant correlation between serum fluoride and age. No sex related difference was found.

Serum and urine ionic fluoride: normal range in a nonexposed population.

The present study was undertaken to evaluate the fluoride status in the general healthy population of Barcelona. Serum and urine fluoride ionic concentration was determined in a random sample of 250 subjects (age range 15-90 yr) by the Orion fluoride electrode system to determine the normal range of fluoride in this population. The results obtained show that in the general population of Barcelona, fluoride ionic serum concentration ranges between 1 and 47 microg/L (x = 17.5 +/- 9.7 microg/L) and fluoride ionic urine concentration ranges between 156 and 1990 microg/24 h (x = 671 +/- 373 microg/24 h). The mean serum fluoride concentration of the younger population was shown to be significantly greater (p < 0.05) than that of the older group. No sex-related difference was found.

Influence of posttransplant time on dose and concentration of tacrolimus in liver transplant patients.

The dosage of tacrolimus (D), the trough blood concentrations (C) and the evolution of the D/C ratio were followed for 1 year after transplantation in so adult patients (38 males and 12 females) undergoing liver allograft. A total of 1489 samples were analysed by the IMx tacrolimus method. The overall median concentration was 11.27 ng/ml. During the 1st month the median of the tacrolimus levels was 8.4 ng/ml, and 73.1% of the analysed samples were within the established therapeutic range. The median oral tacrolimus dose was progressively reduced from 0.12 mg/kg per day during the 1st month to 0.06 mg/kg per day at the end of studied period. A significant negative association was observed between the D/C ratio and the post-transplantation period (r = -0.3892; P < 0.0001). The median D/C ratio ranged from 0.0144 at 1st month to 0.0053 at 1 year. Significant D/C declines were observed after the 1st and 3rd months posttransplant. The decrease in corticosteroid doses and the increase in serum albumin may explain the reduction in clearance with time.

Glutathione S-transferase M1 (GSTM1) and T1 (GSTT1) polymorphisms and lung cancer risk among Northwestern Mediterraneans.

Several polymorphic genes including those encoding for glutathione S-transferases (GST) have been reported to be involved in modifying lung cancer risk in smokers. The gene GSTM1 is frequently deleted in humans and a possible association between the null genotype and lung cancer risk is controversial. Another polymorphic gene of the same supergene family, GSTT1, is also involved in the detoxification of some environmental carcinogens. Both genes were genotyped in (a) a group of lung cancer patients (n = 160); (b) a group of healthy smokers (n = 120); (c) a group of blood donors from the general population (n = 192). All patients and controls were Northwestern Mediterranean Caucasians. The results show that the GSTM1 null genotype (GSTM1*0/GSTM1*0) was slightly over represented in the lung cancer patients (frequency of 58%; OR: 1.40, 95% CI: 0.74-2.61, referred to healthy smokers). The histological type most clearly modified was small cell carcinoma (frequency of 62.2%, OR: 1.91, CI: 0.78-4.69). The subdivision of the patients with one or two copies of the GSTM1 gene according to a GSTM1*A, GSTM1*B or GSTM1*A/B genotype (frequencies of 28.2%, 11.2%, 2.5% respectively) revealed no significant differences between the cases and both control groups. The frequency of the deleted GSTT1 genotype among the lung cancer patients (24%) was not significantly increased (OR: 1.08, CI: 0.57-2.05, referred to healthy smokers). The results showed that 14.4% of the patients presented homozygous deletion of both GSTT1 and GSTM1 (12.5% among healthy smokers) suggesting no potentiation between null genotypes for lung cancer risk.

Serum selenium concentration of a healthy northwest Spanish population.

Selenium (Se) is an essential element, cofactor for glutathione peroxidase (GSHPx) activity, whose deficiency may induce modifications in the cellular antioxidative status and induce the appearance of different diseases. Current views suggest that a serum Se concentration inferior to 45 micrograms/L may correlate with an increased risk of coronary hearth diseases, coronary atherosclerosis and cancer. Since the Se concentration in human blood varies between geographical areas, we initiated a study to evaluate the Se status in the general healthy population of Barcelona. Serum Se concentration was investigated in a random sample of 150 subjects (age range 18-70 yr) by graphite furnace atomic spectrometry (FLAAS). L'vov platform, Zeeman background correction, and other specifications of stabilized temperature platform furnace (STPF) concept were followed. The results show that in the general population of Barcelona, Se serum concentration ranges between 60 and 106 micrograms/L (X = 80.7 +/- 10 micrograms/L). These values can be considered within the safe limits, since no subject was found with a concentration lower than the threshold of 45 micrograms/L.

[Exposure to lead among the population of Barcelona: chronologic trends from 1984 to 1995]. / Estudio de la exposición al plomo en la población de Barcelona: evolución cronológica entre 1984 y 1995.

BACKGROUND: This study was designed to determine current lead exposure in the Barcelona population and to evaluate the changes occurred during the last 10 years. Blood lead concentration was investigated in a random sample of 694 healthy subjects (age range: 0-65 years). PATIENTS AND METHODS: Adults were random selected from a group of blood donors. Samples of children analysed were selected from subjects with a preoperatory analyses without any disease that could modify blood lead levels. Lead levels were determined by atomic absorption spectrometry. RESULTS: Blood lead concentration was 4.06 +/- 1.4 micrograms/dl in umbilical cord, 8.9 +/- 2.9 micrograms/dl in the paediatric population and 7.8 +/- 4.2 micrograms/dl in the total of adults analyzed. There was statistical differences between the younger subjects and the older population. In 1984 the results found were 18.6 +/- 6.6 micrograms/dl. CONCLUSIONS: The results obtained show that in the last 10 years a reduction on the blood lead levels was occurred. This reduction is parallel with a diminish in the lead petrol concentration in the ambient air.

Metabolism of hexachlorobenzene in humans: association between serum levels and urinary metabolites in a highly exposed population.

Serum and urine from 100 subjects of a general population highly exposed to airborne hexachlorobenzene (HCB) were analyzed to obtain new insights into the metabolism of this ubiquitous compound. HCB was detected in all serum samples with concentrations ranging between 1.1 and 953 ng/ml. The major known metabolites of HCB were investigated in urine collected over 24 hr. Pentachlorophenol (PCP) was detected in all urines with values ranging between 0.58 and 13.9 micrograms excreted in 24 hr [mean +/- standard deviation (SD), 2.52 +/- 2.05; geometric mean, 2.05]. A sulfur derivative that, after hydrolysis, yielded pentachlorobenzenethiol (PCBT) could also be identified and quantified in all the urines with values ranging between 0.18 and 84.0 micrograms of PCBT excreted in 24 hr (mean +/- SD, 3.47 +/- 10.8; geometric mean, 1.39). The sulfur derivative assessed as PCBT appeared to be the main metabolite, with urinary concentrations surpassing those of PCP in the subjects with higher HCB accumulation (HCB in serum > 32 ng/ml). PCBT concentration in urine collected over 24 hr showed a very strong association with HCB concentration in serum; the association was stronger in males than in females. An increase of 1 ng/ml of HCB in serum led to an increase of 2.12 micrograms of PCBT excreted in urine collected over 24 hr in males (95% CI, 1.82-2.44) and to an increase of 0.67 microgram of PCBT in females (CI, 0.33-1.09). A weaker association was found between PCP in urine and HCB in serum, which was only statistically significant in males (an increase of 1 ng/ml of HCB in serum led to an increase of 0.63 microgram of PCP excreted in urine collected over 24 hr; (CI, 0.34-0.95). These results show that the formation of the cysteine conjugate is a quantitatively more important metabolic pathway in humans than the formation of PCP. Moreover, the association found suggests that PCBT is a good urinary marker of HCB internal dose and glutathione-mediated metabolism.

Comparative analysis of tacrolimus (FK506) in whole blood liver transplant recipients by PRO-TRAC enzyme-linked immunosorbent assay and microparticle enzyme immunoassay IMX methods.

The macrolide tacrolimus (FK506) is a powerful immunosuppressive drug that acts early in the T-cell activation process and inhibits cytokine gene transcription. Data from several trials in liver transplantation have shown the efficacy of tacrolimus in the prevention of allograft rejection and its potent hepatotrophic effect, which could explain its great success in liver transplantation. However, tacrolimus is not devoid of adverse effects (mainly nephrotoxicity and neurotoxicity) requiring careful blood level monitoring, which is an essential aid in the adjustment of drug dosing. Several methods of analysis are available to measure tacrolimus in whole blood. A new assay based on the enzyme-linked immunosorbent assay (ELISA) technology has been developed. The INCSTAR PRO-TRAC FK506 is a sensitive immunoassay (range, 0.5 to 60 ng/ml), which uses a mouse monoclonal antibody to FK506. Samples are extracted into methanol and dried under nitrogen. The reconstituted extracts are analyzed by ELISA by using 2-h incubation. The aim of this study was to evaluate the ELISA method in routine monitoring of liver transplant patients and to compare the whole blood results with those obtained by Abbott microparticle enzyme immunoassay (MEIA) IMx. Precision studies with 20 samples from 4.37 and 17.1 ng/ml gave within-run total coefficients of variance of 14.4 and 17.4%, respectively. A total of 63 blood samples was analyzed. The mean +/- SD were 9.68 +/- 5.92 and 10.52 +/- 7.54 ng/ml by ELISA and MEIA assays, respectively. There was an acceptable correlation between the methods: ELISA = 1.419 + 0.785 MEIA; Sy x x = 2.639; r = 0.804. Serial tacrolimus measurements (n = 13) in two patients with bilirubin levels > 20 mg/dl yielded mean +/- SD (range) of 11.64 +/- 7.59 ng/ml (2.60-25.40 ng/ml) and 15.55 +/- 10.78 ng/ml (3.60-34.4 ng/mL) by ELISA and MEIA assays, respectively. These discrepancies in concentrations can result from variation in matrix or different cross-reactivities or both in the two tests. We concluded that the INCSTAR PRO-TRAC FK506 is suitable for routine whole blood tacrolimus monitoring.

Glutathione-S-Transferase M1 and codon 72 p53 polymorphisms in a northwestern Mediterranean population and their relation to lung cancer susceptibility.

Several polymorphic genes have been reported to be possibly involved in modifying lung cancer risk in smokers. The gene GSTM1 is frequently deleted in human populations, and the null genotype has been reported to be a risk factor for developing lung carcinoma. A germline polymorphism of p53 with a single-base change at codon 72 that causes an amino acid replacement of arginine (Arg; CGC) by proline (PRO; CCC) has also been reported to be associated with cancer susceptibility in a Japanese population. Both polymorphisms were genotyped by PCR in a northwestern Mediterranean healthy population (n = 147) and in a group of lung cancer patients (n = 139). The results showed that the frequency of the GSTM1 null genotype was higher in the lung cancer patients compared to the controls [odds ratio (OR), 1.57; 95% confidence interval (CI), 0.99-2.51]. The histological subtypes most clearly modified were small cell carcinoma (OR, 1.89; CI, 0.97-3.65) and adenocarcinoma (OR, 1.93; CI, 0.90-4.14). The null GSTM1 genotype was more frequent among those cancer patients who were medium/ light smokers (< or = 50 pack-years) and in those who showed an onset of the disease at a more advanced age. The study of the p53 polymorphism in the healthy population showed allele frequencies of 0.79 (Arg) and 0.21 (Pro). The frequencies found in the lung cancer patients were statistically similar. Both polymorphisms were studied together, and the relative risk of the combination null GSTM1 and Pro/Pro or Arg/Pro genotypes was calculated taking the combination of GTSM1 + together with Arq/Arg as a baseline. The OR found (1.97; CI, 1.03-3.73) suggests that the Pro allele of the p53 germline polymorphism may slightly increase the risk fo the GSTM1 null genotype among smokers.

Cadmium and zinc relationships in the liver and kidney of humans exposed to environmental cadmium.

Concentrations of cadmium and zinc were determined in the liver and in the kidney (cortex and medulla) of subjects from the general population of Barcelona (Spain) by atomic absorption spectrometry. Tissues were collected from necropsies of 50 selected subjects without any occupational exposure to heavy metals. Cadmium levels calculated on a fresh tissue basis were 14.6 +/- 5.9 micrograms/g (2.4-31) in the kidney cortex, 8.6 +/- 4.3 micrograms/g (1.5-16.7) in the kidney medulla and 0.98 +/- 0.50 micrograms/g (0.32-2.32) in the liver. Zinc concentrations ranged between 18-53 micrograms/g, (mean +/- S.D.: 38.0 +/- 10 micrograms/g) in the kidney cortex, 25.0 +/- 7.7 micrograms/g (12-42 micrograms/g) in the kidney medulla and 41.7 +/- 18.3 micrograms/g (20-84 micrograms/g) in the liver. The aim of the present work was to study the association of cadmium and zinc in the kidney and in the liver of a human population with cadmium accumulation from an environmental origin. The results obtained showed a significant correlation between cadmium and zinc concentration in the liver (r = 0.86, P < 0.001), but not in the kidney.

Effects of long-term antiepileptic therapy on the catabolism of testosterone.

The serum levels of testosterone, sex hormone binding globulin, and free testosterone index were measured in 51 epileptic men (age 18-45) in order to assess the possible effects of antiepileptic drugs on sexual dysfunction. An analytical gas chromatography-mass spectrometry method was developed to assess the urinary excretion of testosterone, epitestosterone, androsterone, etiocholanolone, 11-OH androsterone and 11-OH etiocholanolone and to evaluate if the catabolism of testosterone had been increased. Twenty normal healthy males of similar age, 18-45 years, served as control group. Patients receiving polytherapy (n = 34) or monotherapy with carbamazepine (n = 8) or phenytoin (n = 9) showed higher levels of sex hormone binding globulin and testosterone, and lower levels of free testosterone than did the controls (P < 0.03). Urinary excretion of the metabolites androsterone and 11-OH androsterone was significantly reduced (P < 0.02) in the polytherapy group, while the monotherapy group showed only significant differences (P < 0.02) in the elimination of 11-OH androsterone. Our results suggest that an induction of the hepatic synthesis of sex hormone binding globulin may be the mechanism by which the antiepileptic drugs lower the levels of free testosterone in serum. However, the reduced excretion of androsterone and the normal levels of etiocholanolone show that the antiepileptic drugs do not produce an increase in the main catabolism pathway of testosterone.

Accumulation of hexachlorobenzene in humans: a long standing risk.

1. Hexachlorobenzene (HCB) internal dose in the general population of Barcelona (Spain) was estimated after new indications of the carcinogenicity of this chemical in humans were recently reported. Hospital blood bank facilities and randomly selected volunteers were used for HCB analyses in serum (n = 100) and cerumen (n = 25). Other main organochlorine residues often found in human tissues and blood (pp DDE, beta-HCH,) were also determined. 2. HCB serum levels currently found (Range 0.7-19.7 ng Ml-1; X +/- s.d.: 4.13 +/- 3.61; GM: 3.05) were compared to those found in a similar survey made in 1986 on the same population. The serum HCB levels showed a significant decrease (P < 0.001) when compared to the former results and correlated with age (P < 0.001) suggesting a progressive preponderance of a stable blood-adipose equilibrium with fewer variations due to recent and variable intake of the chemical. 3. Cerumen analyses revealed detectable concentrations of HCB in all samples (Range: 160-4790 ng g-1 in extractable lipid basis) and confirmed the suitability of this matrix to assess the body burden of residues accumulated in adipose and lipid-rich tissues. The set of results shows that, although HCB exposure has been reduced, the overall population under study still accumulates significant amounts of this possible carcinogen.