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Clinical Trial 2006; 27(Suppl 2): 134-137 PubMed PMID: 17159798 Citation  Keywords:  Aged, Cells, Cultured, Combined Modality Therapy, Diabetic Foot:surgery, Humans, Mesenchymal Stem Cell Transplantation:methods, Skin Transplantation:methods, Transplantation, Autologous, Wound Healing,.   

OBJECTIVES: This study was performed to test a new technique for treatment of chronic non-healing wound (diabetic ulcer) using autologous biograft composed of autologous skin fibroblasts on biodegradable collagen membrane (Coladerm) in combination with autologous mesenchymal stem cells (MSC) derived from the patient's bone marrow.r />r /> DESIGN: The bone marrow aspirate of the patient with diabetic foot was applied directly to the wound and injected into the edges of the wound, finally covered with prepared autologous biograft. The patient received two additional treatments with cultured MSC on day 7 and 17.r />r /> RESULTS: The wound showed a steady overall decrease in wound size and an increase in the vascularity of the dermis and in the dermal thickness of the wound bed after 29 days of combined treatment.r />r /> CONCLUSIONS: Closing and healing of the non-healing diabetic ulcer was achieved by using the given combined therapy....

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Journal Article 2006; 27(Suppl 2): 130-133 PubMed PMID: 17159797 Citation  Keywords:  Adult, Chromatography, High Pressure Liquid:methods, Dose-Response Relationship, Drug, Female, Humans, Male, Methadone:analogs & derivatives, Molecular Conformation, Opioid-Related Disorders:blood, Sensitivity and Specificity,.   

OBJECTIVES: A stereoselective HPLC method was developed to separate and quantify both enantiomers of methadone and its main metabolite EDDP in serum and urine. The method was used to establish that there is a relationship between the dose of methadone prescribed and its serum concentration as well as urine excretion of methadone and its metabolite enantiomers.r />r /> METHODS: The chiral alpha1-glycoprotein stationary phase was used for enantioseparation of (R)-methadone, (S)-methadone and (R)-EDDP (S)-EDDP. The enantiomers of methadone and EDDP were extracted from urine and serum by a simple solidphase procedure.r />r /> RESULTS: The validated method was applied to the analysis of 31 serum and urine samples obtained from methadone-maintained outpatients (65% male, age 28.8+/-4; methadone dose 146+/-47 mg). A significant correlation (Pearson) r=0.67 (p<0.001) between methadone dose and serum concentration of (R)-methadone was found. Due to the large variation in results obtained from analysis of the subjects' urine specimens, no statistically significant relationship between methadone dose and urine excretion of methadone and EDDP enantiomers was established. The rate of R/S methadone (1.38 in serum, 2.43 in urine) and R/S EDDP (0.83 in urine) confirmed stereoselectivity in methadone metabolism with high individual variability.r />r /> CONCLUSIONS: The enantioselective evaluation of serum methadone concentration might be an interesting tool in methadone maintenance programme. On the other hand, the urinary excretion of methadone and EDDP enantiomers is not reliable as marker of methadone compliance but could be useful for monitoring individual metabolism or for studying the stereoselectivity in pharmacokinetics and metabolism of methadone....

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Evaluation Studies 2006; 27(Suppl 2): 125-129 PubMed PMID: 17159796 Citation  Keywords:  Chromatography, Thin Layer, Forensic Toxicology:methods, Gas Chromatography-Mass Spectrometry, Hallucinogens:blood, Humans, Models, Biological, Substance Abuse Detection:methods,.   

OBJECTIVES: Until recently, routine toxicological analysis of some hallucinogens in biological material posed problems which were only resolved after the introduction of modern analytic systems into toxicological laboratories. The most frequent hallucinogens in clinical and forensic toxicology can be grouped as: cannabinoids, tropane alkaloids, N,N-dimethyltryptamine derivatives and synthetic or semisynthetic hallucinogens.r />r /> METHODS & RESULTS: There are several methods currently used for their analysis. Immunoassay analysis of abused hallucinogens is limited to the cannabinoids. Thin layer chromatography (TLC) is able to detect higher concentrations of 1-nor-delta- 9-tetrahydrocannabinol-9-carboxylic acid (THC-COOH), tropane alkaloids (atropine and scopolamine) and ketamine (synthetic hallucinogen) in urine but for lower concentrations and for some other substances it lacks sensitivity. A reliable solution to the demand for specific and sensitive analysis of hallucinogens in biological material is gas chromatography - mass spectrometry (GC-MS). Thus, at present, analysis of cannabinoids, tropane alkaloids, ketamine as well as psilocin (N,N-dimethyltryptamine derivative) is well-managed.r />r /> CONCLUSIONS: The introduction of GC-MS systems appears to be indispensable for satisfactory qualitative and quantitative analysis of drugs of abuse, particularly hallucinogens in biological material....

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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.r />r /> 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.r />r /> 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.r />r /> 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.r />r /> 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.r />r /> 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.r />r /> 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.r />r /> 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.r />r /> 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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Evaluation Studies 2006; 27(Suppl 2): 108-111 PubMed PMID: 17159792 Citation  Keywords:  2-Methyl-4-chlorophenoxyacetic Acid:toxicity, Alanine Transaminase:blood, Alkaline Phosphatase:blood, Animals, Creatinine:blood, Dogs, Female, Herbicides:toxicity, Male, Organ Size:drug effects, Spermatogenesis:drug effects, Testis:anatomy & histology, To.   

OBJECTIVES: This study was performed to investigate the toxicity of the herbicide MCPA after repeated treatment according to OECD No 409.r />r /> METHODS: The herbicide MCPA was administrated to dogs of both sexes in daily doses: 0, 1, 5 and 15 mg/kg per os during 90 days. Clinical observations were recorded, opthalmoscopic, haematological, clinical chemistry parameters were investigated. The morphological changes were examined using pathological and histological techniques.r />r /> RESULTS: An increase in the activity of the enzyme ALT and of urea concentration was observed in the middle- and high-dose groups. ALP and creatinine were decreased in dogs of all dose groups. After 60 days of MCPA administration, the concentration of glucose decreased in the high-dose group. The relative weight of both testes was decreased in the males from the high-dose group in comparison to controls. The decrease in testes weight was associated with focal testicular atrophy and loss of spermatogenic cells in the lobuli of testes. Focal hepatocellular necrosis, inflammatory changes and mononuclear nodules were observed in the liver, predominantly in males of the high-dose group.r />r /> CONCLUSIONS: Administration of MCPA did not cause mortality and had no adverse effects on hematological parameters of animals. The results of the study showed that MCPA treatment caused inflammatory lesions in the liver, moderate damage of the kidney, and had focal adverse effect on the development of spermiogenesis. The changes were reversible....

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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).r />r /> 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.r />r /> 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).r />r /> 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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