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Journal Article 2009; 30(Suppl 1): 219-224 PubMed PMID: 20027174 Citation  Keywords:  Cholesterol:blood, Chronic Disease:epidemiology, Environmental Pollutants:blood, Follow-Up Studies, Humans, Male, Middle Aged, Occupational Exposure, Polychlorinated Dibenzodioxins:blood, Prevalence, Triglycerides:blood,.   

OBJECTIVES: The aim of this study, performed in 2008, was to evaluate the consequences of severe occupational intoxication with 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD) that occurred during production of the herbicide trichlorophenoxyacetic acid in the period 1965-1968.

DESIGN: Examination of 11 men, mean age 64.4+/-1.5 years, included: internal and neurological examination, eye fundus examination, TCDD in plasma, thyroid-stimulating hormone (TSH), testosterone and serum lipids, ultrasonography of the carotid artery, nerve conduction study (NCS), electroencephalography (EEG), visual evoked potential (VEP), Lanthony test of acquired visual impairment, single photon emission computer tomography (SPECT) of the brain, neuropsychological examination and carbohydrate-deficient transferrin (CDT), a marker of chronic ethanol intake.

RESULTS: Mean TCDD level in 2008 was still 274.0+/-181.2 pg/g blood lipids (reference level is 2-3 pg/g). All (100%) patients had residues of chloracne/chloracne consequences, atherosclerotic changes on the eye fundus and plaques in the carotid arteries. Progression of intima-media thickness (IMT) from a mean of 0.84+/-0.14 mm in 2003 to 1.09+/-0.18 mm in 2008 was observed. Ninety-one per cents of patients had impairment in SPECT of the brain; and 55% of patients had hyperfixation of the perfusion indicator as a measure of increased neuronal activity. Additionally, 91 % of patients were treated for hyperlipidaemia, 73 % for hypertension, 55 % for diabetes type 2, 45 % for ischemic heart disease, and 36 % for psychological disorders. The Lanthony test demonstrated acquired dyschromatopsia in 80 % of patients. Mean colour confusion index (CCI) was 1.438, which indicates impairment since 2003, when the index was 1.302. CDT was in the normal range and did not correlate with CCI. Neuropsychological status appeared stabilized in all 8 patients examined, with impairment in one or more parameter (memory, attention, verbal fluency, psychomotor speed, motorics) in comparison to the norm.

CONCLUSION: Forty years after intoxication, the blood level of TCDD is still 100 times higher than in the general population. Other PCDD/Fs were not elevated. A high percentage of subjects suffer from neurological and vascular disorders. No association of alcohol consumption with neurological impairment was seen, and the highly significant correlation between CCI and TCDD blood concentration suggests that acquired colour impairment was associated with TCDD but not with alcohol consumption. IMT significantly increased during past 5 years. The patients obviously need complex treatment, including intense hypolipidaemic and antidepressant therapy....

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Journal Article 2009; 30(Suppl 1): 182-185 PubMed PMID: 20027168 Citation  Keywords:  Age of Onset, Aged, Carcinogens, Environmental:toxicity, Case-Control Studies, Chromium:toxicity, Humans, Lung Neoplasms:epidemiology, Male, Middle Aged, Occupational Exposure, Small Cell Lung Carcinoma:epidemiology, Time Factors,.   

OBJECTIVE: Workers chronically exposed to hexavalent chromium have higher incidence of lung cancer. Our study investigates incidence of lung cancer types, age at onset of the disease and survival time among chromium exposed workers (smelters, tapers, crane operators) in comparison to non-exposed persons.

METHODS: 64 chromium exposed workers and 104 male controls with diagnosed lung cancer were analysed. The average exposure time among workers was 16.71 +/- 10.02 (S.D.) years (range 1- 41 years).

RESULTS: Chromium exposure significantly decreases the age at the onset of the disease by 3.51 years (62.20 +/- 9.08 years in exposed group and 65.71 +/- 10.50 years in control; P=0.018). Small cell lung carcinoma (SCLC) forms 25.0 % of all cases in chromium exposed workers and 16.34% in non exposed individuals. No correlation was found between the age at the diseases onset and time of exposure. The mean survival time in exposed group was 9.03 +/- 12.73 month, in control 12.14 +/- 21.94 month, but this difference was not significant (P=0.473).

CONCLUSION: Occupational exposure to chromium was identified as an important risk factor of lung cancer, decreasing the age at the diseases onset. Higher percentage of SCLC was found in chromium exposed individuals....

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Journal Article 2009; 30(Suppl 1): 177-181 PubMed PMID: 20027167 Citation  Keywords:  Adolescent, Adult, Aged, Aging, Animals, Child, Child, Preschool, Dental Amalgam, Diet, Environmental Exposure:analysis, Female, Fishes, Hair:chemistry, Humans, Male, Mercury:analysis, Middle Aged, Sex Characteristics, Young Adult,.   

OBJECTIVES: Exposure to mercury, a risk factor for neuro-developmental toxicity, was evaluated in the Czech Republic by performing mercury determination using human hair as an indicator.

METHODS: Hair samples from Czechs (n=311; 2-66 years old) were analyzed for mercury content. Total mercury was analysed by mercury analyzer AMA 254.

RESULTS: The highest total mercury content found in sampled hair was 3.55 microg/g and the lowest content was 0.015 microg/g. No correlation was found between the mercury levels in the sampled hair and the subject's age, gender, and the amount of amalgam fillings. A total of 38 hair samples were analyzed for methylmercury content.

CONCLUSION: The results show a positive correlation between the total mercury content in human hair and the consumption of marine and freshwater fish. Hair are a very good indicator of fish consumption....

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Journal Article 2009; 30(Suppl 1): 156-162 PubMed PMID: 20027164 Citation  Keywords:  Animals, Capsules, Chromatography, Gas, Cod Liver Oil:analysis, Dietary Supplements:analysis, Fish Oils:analysis, Food Contamination, Gadiformes, Gas Chromatography-Mass Spectrometry, Humans, Liver:chemistry, Mercury:analysis, Methylmercury Compounds:anal.   

OBJECTIVES: The aim of this study is to determine the content of mercury, methylmercury and persistent organic pollutants in the capsules filled with fish oil from marine fish and in the canned cod liver and find out, whether analyse products are safe and suitable for human consumption.

METHODS: Total mercury was determined by cold vapour atomic absorption spectroscopy on an AMA-254 (Altec Ltd., Czech Republic) single-purpose mercury analyzer. Methylmercury in the cod liver in the form of chlorid methylmercury was determined by gas chromatography. Seven indicator congeners of PCB (PCB 28, 52, 101, 118, 138, 153, 180), HCH, HCB, DDT and its degradation products DDE and DDD, were determined by gas chromatography-mass spectrometry.

RESULTS: In capsules filled with fish oil (n=19) total Hg levels were in the range of 0.013 to 2.03 ng/g. All the capsule oil samples analyzed for MeHg were below the detection limit. The highest concentration of total Hg was found in cod liver - can A (0.223 ng/g). The maximum percentage of MeHg in total Hg concentration was found in a liver sample - can B. The values of alpha-, beta-, gamma-, delta- HCH and of HCB were found below the detection limits in all the capsule samples analyzed. In eight capsules, the presence of highly lipophilic PCB congeners was demonstrated. DDT and its important DDE metabolite were found in ten samples. In canned cod liver the highest concentration of all PCB congeners was demonstrated in can D. The lowest congener concentrations, however, were found in smoked cod liver - can C. DDT with its main metabolite DDE was detected in can C. No additional DDT and DDD persistent pollutants were detected.

CONCLUSIONS: The consumption of fish oil in capsules, and canned cod liver is safe and healthy and should be encouraged....

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Journal Article 2009; 30(Suppl 1): 137-142 PubMed PMID: 20027160 Citation  Keywords:  Adult, Alleles, Azathioprine:adverse effects, Female, Genetic Predisposition to Disease, Genotype, Humans, Leukopenia:chemically induced, Male, Methyltransferases:genetics, Polymerase Chain Reaction, Polymorphism, Genetic, Polymorphism, Single Nucleotide,.   

OBJECTIVES: Our study aims to find the relationship between metabolic enzyme thiopurine S-methyltransferase (TPMT) gene polymorphisms and clinical output of the therapy with azathioprine. We focused on patients who experienced leucopenia caused by high blood levels of active azathioprine metabolites.

DESIGN: Our group consists of 87 patients who have been treated by azathioprine. 21 individuals experienced leucopenia during treatment with standard dose of azathioprine. We have used PCR-REA and "real-time" PCR methods for genotype detection G238C, G460G and A719G substitutions in TPMT gene.

RESULTS: We have found statistical association between the presence of non-standard TPMT alleles and adverse event associated with azathioprine treatment - leucopenia (p=0.0033).

CONCLUSION: Our results confirm that TPMT genotyping prior to the treatment with azathioprine could predict patients with genetic predisposition for serious leucopenia and seems to be a useful genetic marker for individualisation of the therapy....

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Comparative Study 2009; 30(Suppl 1): 133-136 PubMed PMID: 20027159 Citation  Keywords:  Benzothiepins:pharmacology, Brompheniramine:pharmacology, Chlorpheniramine:pharmacology, Dose-Response Relationship, Drug, Extracellular Space:drug effects, Histamine H1 Antagonists:administration & dosage, Humans, Intracellular Space:drug effects, Lorata.   

OBJECTIVES: We analysed and compared the effect of five H1-antihistamines on stimulated oxidative burst at extra- and intracellular level of isolated and stimulated human polymorphonuclear leukocytes.

DESIGN: Oxidative burst of isolated human neutrophils was studied by means of luminol and isoluminol enhanced chemiluminescence.

RESULTS: The following rank order of potency for H1-antihistamines to decrease chemiluminescence was evaluated extracellularly: dithiaden> loratadine> chlorpheniramine> brompheniramine> pheniramine and at intracellular site: loratadine> dithiaden.

CONCLUSION: H1-antihistamines differ substantially according to their chemical structure in suppressing oxidative burst both at extra- and intracellular site of isolated stimulated human neutrophils....

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Journal Article 2009; 30(Suppl 1): 128-132 PubMed PMID: 20027158 Citation  Keywords:  Absorption, Animals, Cattle, Centrifugation, Collagen Type I:chemistry, Electrophoresis, Polyacrylamide Gel, Humans, Hydrogen Peroxide:chemistry, Hydroxyl Radical:chemistry, Luminescence, Oxidation-Reduction, Phagocytes:metabolism, Platelet Aggregation, P.   

OBJECTIVES: The effects of non-modified and oxidatively modified calf skin collagen type I on platelet aggregation and the oxidative burst of phagocytes were examined in the framework of a general hypothesis that collagen, platelets and phagocytes cooperate to modulate the oxidative burst of phagocytes and the extent of oxidative stress.

MATERIALS AND METHODS: Calf skin collagen type I was subjected to oxidative modification by hydrogen peroxide or hydroxyl radical. Thermal denaturation of collagen was performed in a spectrophotometer equipped with a temperature gradient device. The aggregation of isolated human platelets obtained after differential centrifugation was measured using a dual-channel aggregometer. The production of reactive oxygen species by human whole blood phagocytes was evaluated by luminol-enhanced chemiluminescence.

RESULTS: Oxidative modification of collagen samples was characterized by a decrease in denaturation transition temperature. Oxidatively modified samples showed a modified SDS-PAGE pattern, evidencing a significant destruction of the collagen. All oxidatively modified collagen samples, independent of the oxidation treatment applied, lost their platelet-aggregating and phagocyte oxidative burst-inducing activity.

CONCLUSION: The results suggest that reactive oxygen species were able to modify collagen. On the other hand, oxidatively modified collagen lost its activating properties towards platelets and phagocytes....

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Comparative Study 2009; 30(Suppl 1): 121-127 PubMed PMID: 20027157 Citation  Keywords:  3T3 Cells, Adenosine Triphosphate:metabolism, Animals, Cell Count:methods, Cell Line, Cell Proliferation, Cells, Cultured, Colorimetry, Dermis:cytology, Electric Impedance, Fibroblasts:cytology, Humans, Keratinocytes:cytology, Luminescent Measurements, Mi.   

OBJECTIVES: Determination of cell numbers is a crucial step in studies focused on cytokinetics and cell toxicity. The impedance-based analysis employing electronic sensor array system xCELLigence System allowing label-free dynamic monitoring of relative viable adherent cell amounts was compared with the most utilized methods for relative quantification of viable cell numbers based on a determination of cellular metabolism.

DESIGN: Colorimetric assay based on reduction of tetrazolium salt (MTT) by mitochondrial enzymes and chemiluminiscent assay based on intracellular adenosine triphosphate (ATP) determination were compared with the impedance-based system. Cell morphology was compared by microscopic evaluation. Normal human epidermal keratinocytes (NHEK) and normal human dermal fibroblasts (NHDF), together with 3T3 mouse fibroblast and HaCaT keratinocyte cell lines were employed.

RESULTS: The progress of cell growth curves obtained by different methods during 72 hours reflected cell type and cell seeding densities. The impedance-based method was found to be applicable for the determination of the cell proliferation of 3T3 fibroblasts, HaCaT and NHDF, since the comparison of this method with ATP and MTT determinations showed a comparable results. In contrast, the proliferation of NHEK measured by the impedance-based method did not correlate with other methodological approaches. This could be accounted to the specific morphological appearance of these cells.

CONCLUSION: The study shows the impedance-based detection of viable adherent cells is a valuable approach for cytokinetics and pharmacological studies. However, the specific morphological characteristics of cell lines have to be considered employing this method for determination of cell proliferation without using other reference methods....

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