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Evaluation Studies 2006; 27(Suppl 2): 121-124 PubMed PMID: 17159795 Citation  Keywords:  Amphetamines:urine, Designer Drugs:analysis, Gas Chromatography-Mass Spectrometry:methods, Humans, Reproducibility of Results, Sensitivity and Specificity, Substance Abuse Detection:methods,.   

OBJECTIVES: The purpose of this study is to evaluate the gas chromatographicmass spectrometric method (GC-MS) for assay of stimulants of amphetamine type: amphetamine (AMP), methamphetamine (MAMP), ephedrine (EPH), norephedrine (NOREPH), and the amphetamine-derived designer drugs 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA, ecstasy), 3,4-methylenedioxy-N-ethylamphetamine (MDEA) and N-methyl-benzodioxolylbutanamine (MBDB) in urine. These drugs are often encountered in forensic and clinical toxicological analysis.

METHODS: GC-MS method after mixed-mode solid-phase extraction (SPE) and derivatization with heptafluorobutyric anhydride (HFBA) is presented for quick and reliable screening as well as identification and quantification of amphetamines and amphetamine derived designer drugs in urine.

RESULTS: The measurement of calibration dependence allowed to determine the extent of linearity in the concentration range from 10 to 2,000 ng/ml for all amphetamines except AMP and for all amphetamine-derived designer drugs with correlation coefficients exceeding 0.98. Detection limits were established at 5 ng/ml and quantification limits at 10 ng/ml for all analytes, except AMP. LOD for AMP was established at 20 ng/ml and LOQ at 50 ng/ml. Extraction efficiency for all analytes at concentration levels 50 and 500 ng/ml (n=6) was established in the range 60-99%. Repeatability was measured at concentration levels 50 and 500 ng/ml (n=6), relative standard deviations (RSDs) were under 10% for all analytes.

CONCLUSIONS: The GC-MS method after mix-mode solid-phase extraction and derivatization with heptafluorobutyric anhydride is presented for screening followed by identification and quantification of AMP, MAMP, EPH, NOREPH, MDA, MDMA, MDEA, and MBDB. The applicability of the GC-MS method was proven by analyzing authentic urine samples....

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Journal Article 2006; 27(Suppl 2): 116-120 PubMed PMID: 17159794 Citation  Keywords:  Brain Injuries:blood, Gene Expression Profiling, Humans, Magnetic Resonance Imaging, Metallothionein:blood, Nerve Growth Factors:blood, Phosphopyruvate Hydratase:blood, Radiography, S100 Calcium Binding Protein beta Subunit, S100 Proteins:blood,.   

OBJECTIVES: The aim of this study was to evaluate the correlation of neuron specific enolase (NSE), protein S100B and time-profile of Glasgow Coma Score (GCS) development with metallothionein (MT) blood levels in patients with traumatic brain injury (TBI) during 10 days of hospitalization. Patients were divided into 2 groups with respect to NSE and S100B levels - with (group I) and without (group II) GCS improvement.

METHODS: Serum NSE and S100B concentrations were measured by immunochemical methods; serum metallothionein concentration by electrochemical technique. Cortical biopsies were investigated immunohistochemically and by electron microscope. A cDNA microarray containing 700 gene probes was used to study the changes in gene expression in the ipsilateral cortex.

RESULTS: Values of MT in the blood of group I showed a non-significant decrease compared to group II during 1-3 days after admission. There was an increase of MT during 4-8 days in comparison with values of 1-3 days. The highest value of MT during hospitalization was found in a patient with diffuse axonal injury (group II). The data of cDNA microarray suggested an increase in expression of gene transcripts for oxygen free radical scavenger proteins corresponding with the increase of MT during 4-8 days in both groups.

CONCLUSIONS: The experimental data indicate that monitoring the content of MT in patients with trauma brain injury would be a suitable approach to evaluate the degree of injury or duration of prolonging unconsciousness, particularly in diagnosis of diffuse axonal injury....

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Comparative Study 2006; 27(Suppl 2): 112-115 PubMed PMID: 17159793 Citation  Keywords:  Adult, Aged, Aged, 80 and over, DNA Damage, Diet, Female, Fruit, Humans, Male, Malondialdehyde:blood, Middle Aged, Mineral Fibers:toxicity, Occupational Diseases:prevention & control, Oxidative Stress, Vegetables, Vitamins:blood,.   

OBJECTIVES: In humans, epidemiological evidence suggests that increased consumption of fruits and vegetables can substantially enhance the protection against many common types of cancer.

METHODS & RESULTS: A molecular epidemiological study in 3 Slovak factories producing asbestos, glass fibres and rockwool was conducted. Altogether 388 subjects (239 exposed, 148 controls) were investigated. Food frequency questionnaire was used to ascertain nutrient intake and compared to plasma levels of selected micronutrients, as well as to markers of oxidative stress (MDA, oxidative DNA damage and DNA repair) and antioxidant protection. We found a negative correlation between MDA concentrations and consumption of fruits (p=0.05) and vegetables (p=0.05) in all control subjects. Intake of fruits (p=0.05), vegetables (p=0.01), milk (p=0.01) and cereals (p=0.05) inversely correlated with oxidative DNA damage (net FPG) in all subjects investigated. There was a negative correlation between the intake of fruits (p=0.05) and vegetables (p=0.01) in all exposed subjects.

CONCLUSIONS: Our results suggest that well balanced food consumption with higher fruits and vegetables intake has a protective effect against oxidative damage....

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Comparative Study 2006; 27(Suppl 2): 104-107 PubMed PMID: 17159791 Citation  Keywords:  Animals, Estrogens:toxicity, Parabens:toxicity, Preservatives, Pharmaceutical:toxicity, Toxicity Tests, Vitellogenins:analysis, Zebrafish,.   

OBJECTIVES: Xenoestrogenic potential of propylparaben (PP), one of the most commonly used preservatives in drugs, cosmetics and food, was investigated in vivo using zebrafish (Danio rerio).

METHODS: Juvenile zebrafish (20 days post hatching) were exposed to three different concentrations of propylparaben (PP) dissolved in ethanol and added into the water. After 20 days of exposure the fish were euthanized and vitellogenin concentrations in their whole body homogenates were measured by the method of direct sandwich ELISA. Simultaneously, vitellogenin concentrations in either fish from the control group (exposed to solvent without the substance tested) and in fish from the positive control group (exposed to natural estrogen 17beta-estradiol) were measured.

RESULTS: Vitellogenin concentration in whole body homogenates of control fish was 400 (396-540) ng/ml(-1) (geometric mean (95% CI)). Zebrafish exposure to propylparaben at the concentrations of 0.1; 0.4 and 0.9 mg/l(-1) elicited statistically significant decline (P<0.001) of vitellogenin production, i.e. geometric means of vitellogenin concentrations in whole body homogenates were 240 (186-311); 218 (175-270) and 270 (234-311) ng/ml(-1), respectively. Conversely, the geometric mean of vitellogenin concentration in whole body homogenates of zebrafish exposed to 100 ng/ml(-1) of 17beta-estradiol (positive control) was significantly higher (P<0.001) than values in all other groups, i.e. 35,553 (16,860-74,968) ng/ml(-1).

CONCLUSIONS: Our results suggest an antiestrogenic potential of propylparaben tested in vivo in juvenile zebrafish (Danio rerio). The estrogenic effect of 17beta-estradiol was confirmed....

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Comparative Study 2006; 27(Suppl 2): 100-103 PubMed PMID: 17159790 Citation  Keywords:  Asbestos:toxicity, Asbestosis:genetics, Epoxide Hydrolases:genetics, Gene Frequency, Genetic Predisposition to Disease, Glutathione S-Transferase pi:genetics, Glutathione Transferase:genetics, Health, Humans, Micronuclei, Chromosome-Defective:chemically i.   

OBJECTIVES: As asbestos presents a direct genetic hazard for humans, a small-scale molecular epidemiological study was conducted to monitor 61 subjects long-term exposed to asbestos in comparison with 49 town controls and 21 control subjects from administration of the same factory.

RESULTS: Asbestos exposed workers had significantly higher numbers of chromosomal aberrations compared with both control groups (P=0.003). Clinical examination showed that 44.3% of exposed workers developed symptoms of asbestosis. We were interested in the relationship between the risk of asbestos-coupled diseases and individual variability in biotransformation enzymes, especially in glutathione S-transferases and microsomal epoxide hydrolase. GSTP1*105Val allele appeared less in the group of workers with asbestosis compared to those without asbestosis (18.5% vs 34.7%, P=0.044), and in subjects with developed asbestosis coupled with bronchitis compared to those without bronchitis (0% vs 25%, P=0.048). Similarly, the genotype corresponding to low activity of microsomal epoxide hydrolase was significantly decreased in workers with fibrotic plaques compared to those without plaques (26.7% vs 56.3%, P=0.045).

CONCLUSIONS: Our results suggest that GSTP1*105Val allele and low EPHX1 activity genotype may be protective for people occupationally exposed to asbestos. However, more extensive studies are needed to confirm our results....

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Evaluation Studies 2006; 27(Suppl 2): 82-85 PubMed PMID: 17159786 Citation  Keywords:  Animals, Animals, Newborn, Antioxidants:therapeutic use, Behavior, Animal:drug effects, Carbolines:therapeutic use, Female, Growth and Development:drug effects, Hypoxia:pathology, Male, Motor Activity:drug effects, Pregnancy, Pregnancy, Animal, Premedicat.   

OBJECTIVES: The aim of this study was to evaluate the effect of the synthetic pyridoindole antioxidant stobadine (STO, 50mg/kg/day) on behavioural alterations induced by acute neonatal anoxia in rat offspring.

METHODS: STO was administered orally from day 15 of gestation up to day 7 post partum (PP). Pups of both genders were exposed to anoxia (100% N(2) for 10 minutes in a special glass chamber) on day 6 PP. After the anoxic insult, neurobehavioural development of rats (somatic growth and maturation, neuromotor and reflex development, spontaneous behaviour in open field) was evaluated up to adulthood.

RESULTS: Neonatal anoxia did not affect somatic growth and maturation. The anoxic insult resulted in hyperactivity of male offspring in the open field test. STO pretreatment was found to decrease the anoxia induced hyperactivity in male offspring, and affected also the performance of pups on rotating rod.

CONCLUSIONS: Since STO reduced anoxia-induced hyperactivity, we suggest its potential protective effect in acute hypoxic insults in the perinatal period....

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Comparative Study 2006; 27(Suppl 2): 74-77 PubMed PMID: 17159784 Citation  Keywords:  Anticonvulsants:adverse effects, Congenital Abnormalities:epidemiology, Czech Republic, Developmental Disabilities:chemically induced, Epilepsy:complications, Female, Fetal Death:chemically induced, Humans, Infant, Newborn, Information Services, Pregnancy.   

OBJECTIVES: Care, treatment and follow-up in psychiatric and epileptic pregnant women were compared with women inquiring Czech Teratology Information Service (CZTIS) due to other exposure to drugs during pregnancy.

METHODS: Data were collected by CZTIS, member of European Network of Teratology Information Services from 1996. Exposed groups were compared with pregnant women exposed to drugs which were not classified as major teratogens or hyperthermia. Groups do not vary in age, reproductive history and other parameters.

RESULTS: We observed higher frequency of miscarriage and voluntary termination of pregnancy in the group of psychiatric patients. The number of malformation in prospective follow-up cases was lower than in control group.

CONCLUSIONS: Chronic diseases as epilepsy or psychiatric disorders have to be treated during pregnancy. Women should obtain accurate information about possible risk before pregnancy. Co-operation is needed in these cases. Physicians should keep in mind that appropriate information is to be given to the patient according to her disease, education and comprehension of the problem. If there is any doubt they should organize help for their patients....

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Evaluation Studies 2006; 27(Suppl 2): 69-73 PubMed PMID: 17159783 Citation  Keywords:  Animals, Animals, Newborn, Dietary Supplements, Female, Fetal Hypoxia:chemically induced, Fetal Viability:drug effects, Growth and Development:drug effects, Male, Phenytoin:toxicity, Pregnancy, Pregnancy, Animal, Prenatal Exposure Delayed Effects:preventi.   

OBJECTIVES: The aim of this study was to test the effect of supranutritional dosage of the natural antioxidant vitamin E (VitE) on phenytoin (PHT) induced developmental toxicity and possible long-term effects in rat offspring.

METHODS: PHT (150 mg/kg) was administered by oral gavage daily from day 7 to 18 of gestation and VitE prior to PHT orally on the same days.

RESULTS: PHT administration alone resulted in decreased survival rate and lower body weight of pups on day 21 post partum (PP). Moreover, PHT slightly changed somatic growth and pups failed to present dynamic air righting on days 15-20 PP. VitE supplementation did not alleviate these changes but rather induced persisting body weight reduction on the days 21 PP and 100 PP. We observed also decreased brain wet weight in the PHT and VitE + PHT groups compared to controls.

CONCLUSIONS: We concluded that prenatal supplementation with 500 mg/kg of VitE did not ameliorate the developmental toxicity of PHT and failed to protect postnatal development of rat offspring. Further, in the group supplemented with VitE, the occurrence of persistent body weight gain depression up to adulthood indicates its possible interference with somatic growth regulation....

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