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Case Reports 2011; 32(6): 748-750 PubMed PMID: 22286782 Citation  Keywords:  Adolescent, Anorexia Nervosa:epidemiology, Blindness:epidemiology, Body Image, Comorbidity, Female, Humans, Psychotherapy,.   

: Eating disorders represent significant psychopathological spectrum, particularly in adolescent girls. One of the risk factors associated with eating disorders is a strong emphasis of western visual-oriented culture on physical appearance and slimness. However, sensory handicap leading to disability to see one's own body and bodies of others does not mean that eating disorder cannot develop in blind people. The authors believe that body image is a mental construct that is not dependent on sensory perception only. They present the case of an adolescent patient, blind from early childhood, who shows symptoms of mental anorexia, depression, suicidal behavior and self-harming....

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Journal Article 2011; 32(6): 841-846 PubMed PMID: 22286781 Citation  Keywords:  Adrenergic Agents:pharmacology, Animals, Arachidonic Acids:pharmacology, Behavior, Animal:drug effects, Locomotion:drug effects, Male, Mesencephalon:cytology, Methamphetamine:pharmacology, Mice, Neurons:drug effects, RNA, Messenger:metabolism, Random Allo.   

OBJECTIVES: Since among others also our previous studies suggested an interaction between the endocannabinoid system and methamphetamine brain mechanisms we focused on possible changes in relative expression of cannabinoid CB1 receptor mRNA in mesencephalon from mice sensitized by repeated treatments to methamphetamine stimulatory effects and cross-sensitized by cannabinoid CB1 receptor agonist methanandamide pre-treatment.

METHODS: The Open Field Test was used to measure changes in terms of behavioural sensitization or cross-sensitization to drug effects on locomotion in male mice treated repeatedly with either methamphetamine or methamphetamine after pre-treatment with methanandamide. After each measurement one third of animals were sacrificed and the brain was stored. RNA was isolated from the midbrain and used for reverse transcription and subsequent real-time PCR.

RESULTS AND CONCLUSION: The evaluation of behavioural drug effects showed both development of sensitization to methamphetamine stimulatory effects after repeated treatment and cross-sensitization to them by pre-treatment with cannabinoid receptor CB1 agonist methanandamide. Real-time PCR analyses revealed an increase in CB1 receptor mRNA expression after the first dose of methanandamide followed by decrease after the combined treatment with methamphetamine challenge dose. Our findings suggest that particularly repeated pre-treatment with CB1 agonist methanandamide can elicit increase in the mRNA expression level at least in the mouse mesencephalon neurons associated with cross-sensitization to methamphetamine stimulatory effects....

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Case Reports 2011; 32(Suppl 1): 5-7 PubMed PMID: 22167223 Citation  Keywords:  Brain:growth & development, Cardiovascular System:diagnostic imaging, Cerebral Veins:abnormalities, Echoencephalography, Humans, Infant, Newborn, Intracranial Arteriovenous Malformations:diagnosis,.   

OBJECTIVES: Asphyxia of the newborn has a varied etiology. Clinical consequences have a broad spectrum of presentations. Arteriovenous malformation associated with an aneurysm of the Galen vein can be the cause of focal ischemic changes in the nervous parenchyma.

RESULTS: The authors report a case of a term newborn (birth weight 4, 000 grams, Apgar score 7/9). Physical investigation confirmed the presence of a continuous murmur in the area of the anterior fontanelle. Ultrasonic investigation of the brain detected a huge arteriovenous malformation of the Galen vein. Ultrasonic investigation of the heart excluded structural anomaly, but confirmed a huge retrograde flow in the aorta descendens, opened ductus arterious with suspected formation of coarctation of the aorta and dilatation of the vena cava superior.

CONCLUSION: Congenital anomaly of the Galen vein has a negative influence on prenatal and postnatal development of the brain of a newborn. In the case of our patient, it led to rapid severe asphyxiated changes of the brain parenchyma. Diagnosis and management were established, yet endovascular therapy was not indicated in the early neonatal period....

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Journal Article 2011; 32(Suppl 1): 15-24 PubMed PMID: 22167222 Citation  Keywords:  Alanine Transaminase:blood, Albumins:analysis, Animals, Antioxidants:metabolism, Carps:blood, Ceruloplasmin:metabolism, Dose-Response Relationship, Drug, Drug Evaluation, Preclinical, Environmental Monitoring, Herbicides:pharmacology, Metabolome, Oxidatio.   

OBJECTIVES: Effects of the herbicide formulation Click 500 SC (terbuthylazine 500 g/l) on common carp Cyprinus carpio were assessed through biometric, biochemical, haematological and antioxidant indices, induction of xenobiotic metabolizing enzymes and histological examination of selected tissues.

DESIGN: The fish were exposed to the formulation with terbuthylazine concentrations of 380 ng/l (environmental concentration); 60 µg/l and 550 µg/l for up to 91 days. Haematological indices were assessed using unified methods of haematological examination in fish. Biochemical indices in plasma were measured by biochemical analyzer, ferric reducing ability of plasma (FRAP) and ceruloplasmin activity were determined spectrophotometrically. Concentration of total cytochrome P450, glutathione-S-transferase activity and glutathione content were assessed spectrophotometrically in liver. Activity of liver ethoxyresorufin-O-deethylase (EROD) activity was measured spectrofluorimetrically. Histopathological examination of liver, skin, gills, spleen, cranial and caudal kidney was performed by light microscopy.

RESULTS: An increase (p<0.05) was observed in hepatosomatic index and condition factor in fish from the environmental concentration. A decrease (p<0.05) in haemoglobin and mean corpuscular haemoglobin concentration (MCHC) was found in fish treated with terbuthylazine of 550 µg/l. There was a decline in mean corpuscular volume (MCV) and mean corpuscular haemoglobin (MCH) (p<0.05) in terbuthylazine of 60 µg/l and 550 µg/l. Triglycerides (TAG) (p<0.01) were elevated in all pesticide-treated groups. Alanine aminotransferase (ALT) (p<0.01) and phosphorus (p<0.05) decreased in fish exposed to terbuthylazine of 60 µg/l and 550 µg/l, while albumin (p<0.01) rised in the same groups. An elevation in natrium (p<0.05) in terbuthylazine of 550 µg/l and a rise in protein (p<0.01) in the concentrations of 380 ng/l and 550 µg/l were observed. Correlations between several indices were significant. Ceruloplasmin activity and FRAP were augmented (p<0.01) in the highest concentration tested. Examined xenobiotic detoxification systems were not significantly affected by the exposure. Non-specific histopathological changes were found in the gills and skin of the test fish.

CONCLUSION: The fish treated with terbuthylazine developed a disorder in several haematological and plasma biochemical indices. The levels of markers of oxidative stress increased in response to the exposure. Examined systems involved in detoxification of xenobiotics did not reflect long-term contact with the herbicide. Detected histological lesions were non-specific. The environmental concentration of terbuthylazin affected biometric indices of the test fish....

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Evaluation Studies 2011; 32(Suppl 1): 35-45 PubMed PMID: 22167221 Citation  Keywords:  Administration, Oral, Animals, Animals, Laboratory, Bacterial Toxins:pharmacology, Cyanobacteria:pathogenicity, Eating:physiology, Fish Products:toxicity, Food Contamination, Glutathione:metabolism, Lipid Peroxidation:drug effects, Male, Marine Toxins:pha.   

OBJECTIVES: The toxic cyanobacteria are a serious problem for water supply systems, recreation, and agriculture. Cyanobacteria produce numerous bioactive compounds including microcystins - the most studied cyanobacterial hepatotoxins. Only rare studies addressed realistic situation, i.e. impact of MCs accumulated in the fish tissues on the overall physiology. The aim of the present study was to provide a model simulation of the simple food chain for evaluation of impacts of cyanobacteria on the rat physiology under different exposure scenario.

METHODS: Experimental rats were fed with food with fish meat, which contained external additions of isolated microcystins as well as toxic cyanobacteria Microcystis, nontoxic cyanobacteria Arthrospira and green alga Chlorella. Subgroups of the animals were also challenged with a model antigen KLH to investigated immune-related parameters. We studied parameters of oxidative stress in the liver as levels of lipid peroxidation and glutathion levels. Series of hematological, biochemical and immunological parameters were also investigated.

RESULTS: Although considerable amounts of microcystins were administered to rats, all levels of MCs were under the detection limit (1 ng/g fresh weight) in the rat tissues using tandem LC/MS. Only some conjugates of microcystins with cystein and glutathion were detected in the rat liver exposed to Microcystis biomass (values were around the detection limit). Statistically significant depletion of body and liver weight was observed in groups with microcystin addition in comparison with all other groups. Rats exposed to MCs had stimulated immune system (showed higher antibody answer on administered antigen). Also modulation of some lymphocyte subpopulations was recorded with the most interesting observation of stimulated NK cell numbers in groups exposed to isolated toxins (but not to biomass containing the same toxin amount).

CONCLUSIONS: Our study demonstrates that oral exposure to microcystins in the diet may induce some detoxification responses and modulation of some hematological and immunological parameters....

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Journal Article 2011; 32(Suppl 1): 121-130 PubMed PMID: 22167220 Citation  Keywords:  Animals, Aristolochic Acids:pharmacokinetics, Biotransformation, Carcinogens:pharmacokinetics, Cytochrome P-450 CYP1A1:genetics, Cytochrome P-450 CYP1A2, Cytochrome P-450 Enzyme System:genetics, Cytochromes:genetics, DNA Adducts:metabolism, Drug Evaluatio.   

OBJECTIVE: The herbal drug aristolochic acid (AA) derived from Aristolochia species has been shown to be the cause of aristolochic acid nephropathy (AAN), Balkan endemic nephropathy (BEN) and their urothelial malignancies. One of the common features of AAN and BEN is that not all individuals exposed to AA suffer from nephropathy and tumor development. One cause for these different responses may be individual differences in the activities of the enzymes catalyzing the biotransformation of AA. Thus, the identification of enzymes principally involved in the metabolism of AAI, the major toxic component of AA, and detailed knowledge of their catalytic specificities is of major importance. Therefore, the present study has been designed to evaluate the cytochrome P450 (CYP)-mediated oxidative detoxification and reductive activation of AAI in a rat model.

METHODS: DNA adduct formation was investigated by the nuclease P1 version of the 32P-postlabeling method. The CYP-mediated formation of a detoxication metabolite of AAI, 8-hydroxyaristolochic acid I (AAIa), in vitro in rat hepatic microsomes was determined by HPLC.

RESULTS: Rat hepatic CYPs both detoxicate AAI by its oxidation to AAIa and reductively activate this carcinogen to a cyclic N-acylnitrenium ion forming AAI-DNA adducts in vitro. To define the role of hepatic CYPs in AAI demethylation and activation, the modulation of AAIa and AAI-DNA adduct formation by CYP inducers and selective CYP inhibitors was investigated. Based on these studies, we attribute the major role of CYP1A1 and 1A2 in AAI detoxication by its demethylation to AAIa, and, under hypoxic conditions also to AAI activation to species forming DNA adducts. Using microsomes of Baculovirus transfected insect cells (Supersomes™) containing recombinantly expressed rat CYPs, NADPH:CYP reductase and/or cytochrome b5, a major role of CYP1A1 and 1A2 in both reactions in vitro was confirmed.

CONCLUSION: Based on the results found in this and former studies we propose that AAI activation and detoxication in rats are dictated mainly by AAI binding affinity to CYP1A1/2 or NADPH(P)H:quinone oxidoreductase, by their turnover and by the balance between oxidation and reduction of AAI by CYP1A....

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Journal Article 2011; 32(Suppl 1): 137-145 PubMed PMID: 22167219 Citation  Keywords:  Catalysis, Catechols:metabolism, Chromatography, High Pressure Liquid, Cloning, Molecular, Comamonas testosteroni:enzymology, Electrophoresis, Polyacrylamide Gel, Extracellular Space:enzymology, Hydrogen-Ion Concentration, Mixed Function Oxygenases:geneti.   

OBJECTIVE: Comamonas testosteroni Pb50 is a microorganism that possesses high tolerance for phenol and shows strong phenol degrading activity. This bacterial strain is capable of utilizing phenol as the sole carbon and energy source. Although examples are known in which the C. testosteroni utilizes phenol for growth or metabolism, much less information are known on the nature of the phenol-oxidizing enzymes in this microorganism. Therefore, the occurrence and cellular location of phenol hydroxylase (EC 1.14.13.7), the enzyme participating in the first step of phenol degradation, catalyzing its hydroxylation to catechol in a bacterial Comamonas testosteroni Pb50 strain grown in the presence of phenol as a sole carbon and energy source are the aims of this study.

METHODS: Combination of fractionation with polyethylene glycol 6000 and gel permeation chromatography on columns of Sepharose 4B and Sephacryl S-300 was used for isolation of phenol hydroxylase detectable in the medium in which C. testosteroni was cultivated. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and gel chromatography on Sephacryl S-300 were used to evaluate the molecular mass of the enzyme. The enzyme activity was followed by HPLC (phenol consumption and/or catechol formation).

RESULTS: Whereas low activity of phenol hydroxylase was detected in cytosol isolated from C. testosteroni, more than 16-fold higher activity of this enzyme was found in the medium in which C. testosteroni was cultivated. The presence of phenol hydroxylase extracellular activity suggests that this microorganism may secrete the enzyme into the extracellular medium. Using the procedure consisting of fractionation with polyethylene glycol 6000 and gel permeation chromatography on columns of Sepharose 4B and Sephacryl S-300, the enzyme was isolated from the medium to homogeneity. The formation of catechol mediated by purified phenol hydroxylase is strictly dependent on the presence of NADPH, which indicates that this enzyme is the NADPH-dependent phenol hydroxylase. The enzyme is a homotetramer having a molecular mass of 240 000, consisting of four subunits having a molecular mass of 60 000. The optimum pH of the enzyme for the phenol oxidation is pH 7.6.

CONCLUSION: The results are the first report showing isolation and partial characterization of extracellular NADPH-dependent phenol hydroxylase of a bacterial C. testosteroni Pb50 strain capable of oxidizing phenol to catechol. The data demonstrate the progress in resolving the enzymes responsible for the first step of phenol degradation by bacteria....

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Journal Article 2011; 32(Suppl 1): 95-100 PubMed PMID: 22167216 Citation  Keywords:  Alkaloids:pharmacology, Animals, Antioxidants:metabolism, Dose-Response Relationship, Drug, Drug Evaluation, Preclinical, FMN Reductase:metabolism, Glutathione Reductase:metabolism, Guinea Pigs, Homeostasis:drug effects, Models, Animal, Models, Biological.   

OBJECTIVES: Alzheimer's disease (AD) is a neurodegenerative disorder. Symptomatic treatment is available by inhibitors of acetylcholinesterase (AChE) such as rivastigmine, galantamine and donepezil. As huperzine is a promising compound for AD treatment, our study was aimed at evaluating its pertinent implications in oxidative stress.

METHODS: Laboratory guinea pigs were exposed to huperzine A at doses of 0, 5, 25, 125 and 625 µg/kg. The animals were observed for cognitive disorders and sacrificed one hour after exposure. Tonic-clonic seizures were noticed, but only in highly dosed animals. Ferric reducing antioxidant power (FRAP), thiobarbituric acid reactive substances (TBARS), glutathione reductase and glutathione S-transferase were assessed in frontal, temporal and parietal lobes, the cerebellum, liver, spleen and kidney.

RESULTS: Only minimal changes in enzymatic markers were recognized. Huperzine was not implicated in oxidative stress enhancement as the TBARS values remained quite stable. Surprisingly, antioxidants accumulated in the examined brain compartments as the FRAP value was significantly elevated following all doses of huperzine.

CONCLUSIONS: We discuss the potency of huperzine in enhancing the antioxidant capacity of the central nervous system. Huperzine is probably implicated in more processes than cholinesterase inhibition only....

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