Characterization of drug metabolizing enzymes and assessment of aging in the gilthead seabream (Sparus aurata) liver

Organic anthropogenic compounds such as polycyclic aromatic hydrocarbons, polychlorinated biphenyls, nitroaromatics, dioxins, various pesticides and natural compounds can enter the aquatic environment. These lipophilic compounds are readily taken up into the tissues of aquatic organisms where biotransformation via Phase I and Phase II metabolism can in part, determine the fate and toxicity of the xenobiotics. In this study on gilthead seabream (Sparus aurata), liver microsomal ethoxyresorufin O-deethylase (EROD), methoxyresorufin O-demethylase (MROD), penthyloxyresorufin O-depenthylase (PROD), benzyloxyresorufin O-debenzylase (BROD), aniline 4-hydroxylase (A4H), N-nitrosodimethylamine N-demethylase (NDMA-DE), aminopyrine N-demethylase (APND), caffeine N-demethylase (CN3D) and erythromycin N-demethylase (ERND) were determined. A4H, ERND, NDMA-ND, EROD and PROD activities increased in gilthead seabream (Sparus aurata) liver microsomes of different ages, while CN3D and APND activities decreased. MROD activities were barely detectable in mature fish whereas BROD activities were not detectable at all ages studied. Results were also recorded at the protein level by Western blotting using anti-CYP1A, CYP2E and CYP3A antibodies. In conclusion, elevated levels of gilthead seabream liver microsomal CYP450 enzyme activities might reveal possible exposure to various exogenous compounds, which might affect the desired responses to drugs, hormones and dietary supplements used during breeding. © 2007 Veterinarni Medicina

Natural diterpenoid alysine A isolated from Teucrium alyssifolium exerts antidiabetic effect via enhanced glucose uptake and suppressed glucose absorption

Teucrium species have been used in folk medicine as antidiabetic, antiinflammatory, antiulcer, and antibacterial agents. We have explored in vitro antidiabetic impacts of 2 natural diterpenoids, alysine A and alysine B, isolated from Teucrium alyssifolium. The lactate dehydrogenase (LDH) cytotoxicity assay, glucose uptake test, glucose utilization (glycogen content) test, glucose transport test, glucose absorption (α-glucosidase activity) test, insulin secretion test, RNA isolation and cDNA synthesis assay, qPCR quantification assays, and statistical analyses were carried out in the present study. Alysine A exerted the following effects at non-cytotoxic doses: • Enhanced the glucose uptake, as much as the insulin in the C2C12, HepG2, and 3T3-L1 cells • Increased the glycogen content in the C2C12 and HepG2 liver cells, significantly higher than the insulin and metformin • Suppressed the alpha-glucosidase and the GLUT2 expression levels in the Caco-2 cells • Suppressed the SGLT1 and GLUT1-5 expression levels in the Caco-2 cells • Induced the insulin receptor substrate (IRS)1 and GLUT2 expression levels of the BTC6 pancreatic cells • Induced the insulin receptor (INSR), IRS2, phosphoinositide 3-kinase (PI3K), GLUT4, and protein kinase (PK) expression levels of the 3T3-L1 and C2C12 cells • Increased glucose transport through the Caco-2 cell layer • Did not influence insulin secretion in the pancreatic BTC6 cells Consequently, these data strongly emphasized the antidiabetic action of alysine A on the particularly critical model mechanisms that assume a part in glucose homeostasis, such as glucose uptake, utilization, and storage. Moreover, the expression level of the essential genes in glucose metabolism and insulin signaling was altered in a way that the results would be antihyperglycemic. A blend of in vitro and in situ tests affirmed the antihyperglycemic action of alysine A and its mechanism. Alysine A has exercised significant and positive results on the glucose homeostasis; thus, it is a natural and pleiotropic antidiabetic agent. Advanced in vivo studies are required to clarify the impact of this compound on glucose homeostasis completelyThis work was supported by the Scientific and Technological Research Council of Turkey under TÜBİTAK, under Project No.: 114Z640, and Pamukkale University under Project No.: PAUBAP-2014FBE029

Structural and functional alterations in salivary gland chromosomes and enzyme activity of Chironomus riparius Mg. (Diptera, Chironomidae) from anthropogenically polluted sites in Bulgaria and Turkey

The effect of environment contaminants on genome instability and changes in enzyme activity (acetylcholinesterase (AChE), glutathione S-transferase activities (GST), etoxyresorufin O-deethylase (EROD) and metallothionein (MT)) of Chironomus riparius Mg. from Bulgarian and Turkish stations over two years (2009, 2010) as well as laboratory reared larvae were studied. Physicochemical analysis of the sediments from the field stations indicated the presence of heavy metal pollutants (Cr, Cu, Mn, Pb, and Cd) whose concentrations were higher than the reference data. Genome instability was determined by somatic structural and functional alterations of the polytene chromosomes. In the field sites of both countries somatic aberrations occurred at a significantly higher frequency (p < 0.001) compared with control and laboratory material. C. riparius in sediments with higher concentrations of trace metals (Derincay River, Turkey and Chaya River, Bulgaria), was found to possess a high spectrum of somatic chromosome rearrangements with a somatic index of 2.53 and 3.25 respectively. Changes in functional activity included decreased activity of the Balbiani rings (BRs) and nucleolar organizer (NOR). The observed chromosome alterations agree with the high degree of trace metal pollution and high activity of the studied enzymes. However, no correlation between single somatic chromosome rearrangements and concentrations of specific metal ions was defined. The data are discussed in the light of the wide variety of interactions of metals in nature. The results show that the genome response and biochemical markers are sensitive markers of toxicity and provide early warning indicators of contaminants in the environment. © 2012 Dipartimento di Biologia Evoluzionistica

Yakıotunun Ksenobiyotik Metabolizması Üzerine Etkilerinin ve İlaç-Diyet Etkileşim Potansiyelinin Proteomik ve Moleküler Yaklaşımlar ile Aydınlatılması

TÜBİTAK TBAG Proje15.05.201

Purification and characterization of cytochrome P4501A1 and cytochrome P450 reductase from feral leaping mullet (Liza saliens) liver microsomes

Ph.D. - Doctoral Progra