Azra Bozcaarmutlu

Assessment of pollution in the West Black Sea Coast of Turkey using biomarker responses in fish.

Aim of this study was to determine the extent of pollution in the West Black Sea Coast of Turkey by measuring CYP1A associated EROD activity, phase II enzyme, glutathione S-transferase and antioxidant enzymes, catalase and glutathione reductase activities and immunochemical detection of CYP1A protein level in the liver of mullet. The fish samples were caught from six locations having a varying degree of pollution in the West Black Sea Region of Turkey in August 2005, 2006 and 2007. Mullets caught from Zonguldak Harbour, Ereğli Harbour and Gülüç Streams Mouth displayed 6-9-fold higher EROD, 2-4-fold higher glutathione S-transferase and 2-3-fold higher catalase activities than the reference site, Amasra. Total polyaromatic hydrocarbon levels in mullets caught from these locations were also significantly higher (2-4-fold) than Amasra. The results of this study indicate that Zonguldak Harbour, Ereğli Harbour and Gülüç Stream are highly polluted by polycyclic aromatic hydrocarbons and related contaminants.

Mechanism of Inhibition of CYP1A1 and Glutathione S-Transferase Activities in Fish Liver by Quercetin, Resveratrol, Naringenin, Hesperidin, and Rutin

Quercetin, resveratrol, naringenin, hesperidin, and rutin are phenolic compounds/flavonoids that may have roles in the reduction of cancer susceptibility. In this study, in vitro modulatory effects of them were studied on liver CYP1A1 associated 7-ethoxyresorufin-O-deethylase (EROD) activity and glutathione S-transferase (GST) activity of leaping mullet (Liza saliens). All of the phenolic compounds/flavonoids used exerted an inhibitory effect on both EROD and GST activities of fish. Quercetin, resveratrol, hesperidin, and rutin were found to inhibit EROD activity in a competitive manner; on the other hand, naringenin was found to inhibit EROD activity in a noncompetitive manner. Ki values of quercetin, resveratrol, naringenin, hesperidin, and rutin were calculated from Dixon plots as 0.12 μM, 0.67 μM, 2.63 μM, 18 μM and 0.1 mM, respectively. Resveratrol, quercetin, and hesperidin were found to inhibit GST activity in a competitive manner; on the other hand, rutin and naringenin were found to inhibit GST activity in a mixed-type manner. Ki values of resveratrol, quercetin, hesperidin, naringenin, and rutin were calculated from Dixon plots as 3.2 μM, 12.5 μM, 45 μM, 128 μM, and 150 μM, respectively. The results suggest that quercetin and resveratrol containing foods are effective in the prevention and treatment of cancer.

Combined use of PAH levels and EROD activities in the determination of PAH pollution in flathead mullet (Mugil cephalus) caught from the West Black Sea coast of Turkey

The aim of this study was to determine the extent of polycyclic aromatic hydrocarbon (PAH) pollution by measuring PAH levels and 7-ethoxyresorufin-O-deethylase (EROD) activities in flathead mullet (Mugil cephalus) samples caught from the West Black Sea coast of Turkey. The fish samples were caught in August 2008–2011. The levels of 13 PAHs were measured by high-performance liquid chromatography (HPLC) in the liver of fish. Most of the measured PAHs had three rings (low molecular weight). The frequencies of detection of PAHs were higher in fish samples caught from Zonguldak Harbour and Gülüç Stream Mouth than those from Sakarya River Mouth, Amasra and Kefken. EROD activities and cytochrome P4501A (CYP1A) protein level were also measured in the fish liver microsomes. Highly elevated EROD activities and CYP1A levels were measured in the mullet samples caught from Zonguldak Harbour and Gülüç Stream than those from Amasra and Kefken. The detection of PAHs in the liver of fish samples shows recent exposure to PAHs. The chemical analyses of PAHs and EROD activity results together reflected the extent of PAH pollution in the livers of fish caught from the West Black Sea coast of Turkey. The results indicate that Zonguldak Harbour is the most polluted site in the West Black Sea coast of Turkey.

Aldrin Epoxidation in Flathead Mullet (Mugil cephalus): Possible Involvement of CYP1A and CYP3A

The primary objective of this study was to determine specific cytochrome P450 isozyme(s) involved in the metabolism of aldrin to its toxic metabolite dieldrin in flathead mullet (Mugil cephalus) liver microsomes. To identify the cytochrome P450 isozyme responsible for the aldrin metabolism in mullet liver, the effects of mammalian‐specific cytochrome P450 inhibitors and substrates were determined in the epoxidation reaction of aldrin. CYP3A‐related inhibitors, ketoconazole, SKF‐525A, and cimetidine, inhibited the metabolism of aldrin. The contribution of CYP1A to the aldrin metabolism was shown by the inhibition of 7‐ethoxyresorufin‐O‐deethylase activity in the presence of aldrin. The results indicate that CY1A and CYP3A are the cytochrome P450s involved in aldrin epoxidase activity in mullet. In addition, the suitability of aldrin epoxidase activity for monitoring of environmental pollution was also assessed in the fish samples caught from four different locations of the West Black Sea coast of Turkey.

Purification of CYP2B‐like protein from feral leaping mullet (Liza saliens) liver microsomes and its biocatalytic, molecular, and immunological characterization

In this study, CYP2B‐immunoreactive protein was purified to electrophoretic homogeneity from the liver microsomes of leaping mullet. The purified cytochrome P450 (CYP) gave a single band on sodium dodecyl sulfate–polyacrylamide gel electrophoresis having a Mr of 49,300 Da. Absolute absorption spectrum of the purified CYP showed a maximum at 417 nm and CO‐difference spectrum of dithionite‐reduced cytochrome P450 gave a peak at 450 nm. The purified CYP was found to be active in N‐demethylation of benzphetamine, erythromycin, and ethylmorphine, and O‐dealkylation of pentoxyresorufin in the reconstituted system. However, it was unable to catalyze O‐dealkylation of ethoxyresorufin, methoxyresorufin, benzyloxyresorufin, and hydroxylation of lauric acid and aniline. The purified CYP showed strong cross‐reactivity with anti‐sheep lung CYP2B, a homologue of CYP2B4. N‐terminal amino acid sequence of the mullet P450 had the highest degree of homology with CYP2Bs among the known CYPs. Spectral, electrophoretic, immunochemical, N‐terminal amino acid sequence, and biocatalytic properties of the purified CYP are most similar to those of mammalian cytochrome P4502B. All these data indicate that the purified CYP is certainly 2B‐like. In this study, we not only purified biocatalytically active CYP2B‐like protein from fish, but also demonstrated detailed functional properties of CYP2B‐like protein for the first time.

Modulation of xenobiotic metabolizing enzyme activities in rat liver by co-administration of morin, endosulfan, and 7,12-dimethylbenz[a]anthracene

Abstract Morin is a flavonoid which is present in many plants. Endosulfan and 7,12-dimethylbenz[a]anthracene (DMBA) are toxic chemicals that humans are exposed to in their daily lives. In this study, the protective role of morin was investigated in endosulfan and DMBA treated rats. Eight groups, each comprising seven 2.5-month-old adult male Wistar rats (weighing 170–255 g), were used. Endosulfan, morin, and DMBA were administered individually or in combinations, at 5 mg/kg body weight (bw) (three times/week), 25 mg/kg bw (three times/week), and 30 mg/kg bw (once/week for three weeks) via oral gavage, respectively. On day 54 of the administration period, the rats were killed. DMBA + endosulfan co-administration significantly increased CYP1A1-, CYP1A2-, CYP2E-, and GST-associated activities in the rats compared to the control. DMBA + endosulfan + morin significantly increased CYP1A1, CYP1A2, CYP3A, and GST associated activities in the rats relative to the control. Histopathological studies were performed to investigate protective effects of morin on liver damage. The results indicated that DMBA + endosulfan treatment induced liver damage, and morin reduced this damage. These findings suggest that CYP1A, CYP3A, and GST enzyme activities participate in the protective mechanism of morin against endosulfan and DMBA induced toxicity.