DNA single-strand breakage in rat lung, liver and kidney after single and combined treatments of nickel and cadmium

Single-strand breaks were observed in rat lung and kidney after acute treatment of animals with CdCl2 (4 mg/kg body weight) injected intraperitoneally and NiCl2 (44.4 mg/kg body weight) injected subcutaneously. In the rat liver, no single-strand breakage was evident with those doses in single and combined metal treatments. The most susceptible tissue in rats to cadmium or nickel chloride treatment was the lung tissue. The single-strand breaks were higher in cadmium treatment than in nickel treatment in the rat lung. Also the response to cadmium treatment was obtained earlier than nickel. Rat kidney was also responsive to cadmium treatment. However, the response, although statistically significant, was much lower than the one obtained in rat lung. The combined treatment, which was done by administrating cadmium prior to nickel administration, reduced the number of single-strand breaks significantly and reversed them to control values in rat lung and kidney. This study confirms that cadmium and nickel create single-strand breaks when administered alone in the rat lung. This effect, which was seen in the single metal treatments, was reduced in the combined treatments. (C) 1997 Elsevier Science B.V

COMPARATIVE-STUDIES OF SHEEP LUNG AND LIVER NITROFURANTOIN REDUCTASE

1. Nitrofurantoin reductase which catalyzes the bioactivation of nitrofurantoin was purified to electrophoretic homogenity from sheep liver and lung microsomes, with a yield of 15% and 35%, respectively. The specific activity of both reductases was found to be similar (140 nmol/min/mg protein). 2. The effects of nitrofurantoin and NADPH concentrations, pH, ionic strength, amount of enzyme and reaction period, on the enzyme activity were studied and the optimum conditions for maximum activity of purified fiver and lung nitrofurantoin reductases were determined. 3. The enzyme concentration was found proportional with the square root of the rate of nitrofurantoin reduction up to approximately 15 mug protein/ml and 25 mug protein/ml incubation mixture for liver and lung nitrofurantoin reductases, respectively. 4. The plots of inverse of the nitrofurantoin concentration against the inverse of the square root of the velocity for the reduction of nitrofurantoin by liver and lung enzymes gave K(m) values as 27.78 muM and 32.25 muM, respectively. 5. The purified liver and lung enzymes were also saturated by NADPH at similar concentrations and the K(m) values were calculated as 29.4 muM and 35.5 muM, respectively. 6. The effects of magnesium, nickel, cadmium and copper ions on the nitrofurantoin reductase activity were examined. Magnesium ion was found to have almost no effect, whereas the other ions inhibited the activity of both liver and lung reductases

Adaçayı'nın (salvia Cryptanta) Meme Kanseri Hücre Hatlarında (mcf7 Ve Mda231) İlaç Metabolize Eden Sistemlerin Gen Ekspresyonları Üzerine Etkilerinin İncelenmesi

Önerilen projenin amacı, ülkemizde yetişmekte olan adaçayı (Salvia) türlerinin meme kanser hücrelerine (MCF7 ve MDA231) sitotoksik etkilerinin araştırılması ve bir besin maddesi olarak günlük tüketilen adaçayı ile meme kanseri tedavisinde kullanılan ilaçların olası etkileşiminin moleküler düzeyde incelenmesidir. Bazı ilaçlar vücuda alındığında, hücre içinde, öncelikle karaciğer hücrelerinde, 1. Faz ve 2. faz reaksiyonları ile metabolize edilerek, ilaç olarak etkin ürünlere çevrilir. Ana maddesi etkin bazı ilaçlar ise, aynı sistemler ile etkisizleştirilerek vücuttan atılır. Eş zamanlı olarak tüketilen bitki çayları ile maruz kaldığımız bitkisel kimyasalların, ilaçları metabolize eden enzim sistemleri ile etkileşmesi sonucunda, kullanılan ilacın etkisizleştirilmesi veya vücutta birikim sonucunda toksisitesinin artması mümkündür. Literatürde Salvia ile yapılan çalışmalar bulunmakla birlikte, ilaçları metabolize eden sistemlerin ve bu sistemlerin kontrolünde rol oynayan biyokimyasal yolakların, model olarak seçilen MCF7 ve MDA231 hücrelerinde gen ekspresyonları düzeyinde incelenmesi gereklidir

THE RESPONSES OF HEPATIC MONOOXYGENASES OF GUINEA-PIG TO CADMIUM AND NICKEL

When Cd (3.58 mg CdCl2 . H2O/kg, ip) was administered to male guinea pigs 72 h prior to sacrifice, the metal significantly inhibited the aniline 4-hydroxylase (AH) (16%), ethylmorphone N-demethylase (EMND) (26%), and aminopyrine N-demethylase (AMND) (18%) activities and cytochrome P-450 (12%) and cytochrome b5 (10%) levels. Cd did not alter the hepatic microsomal heme level. Cd, however, significantly increased the hepatic microsomal p-nitroanisole O-demethylase (p-NAOD) (53%) activity. When Ni (59.5 mg NiCl2 . 6H2O/kg, sc) was administered to the guinea pigs 16 h prior to sacrifice, the metal significantly depressed AH (49%), p-NAOD (66%), EMND (47%), and AMND (37%) activities, and cytochrome P-450 (15%), cytochrome b5 (24%), and microsomal heme (28%) levels. For the combined treatment, animals received the single dose of Ni 56 h after the single dose of Cd and then were killed 16 h later. In these animals, significant inhibitions were noted in AH (51%), EMND (47%), and AMND (30%) activities, and cytochrome P-450 (15%), cytochrome b5 (26%), and microsomal heme (30%) compared to those of controls. In the case of p-NAOD activity, the influence was in favor of Ni, i.e., the inhibition was about 61% by the combined treatment. These results reveal that: 1. The response of all substrates of hepatic monooxygenases to Cd are not the same, possibly indicating differential regulation of cytochrome P-450 isozymes by Cd; 2. The inhibitory effect of Ni on hepatic monooxygenases is more profound than that of Cd; and 3. The combination of Cd and Ni does not have a synergistic effect of hepatic monooxygenases of the guinea pig.When Cd (3.58 mg CdCl2 . H2O/kg, ip) was administered to male guinea pigs 72 h prior to sacrifice, the metal significantly inhibited the aniline 4-hydroxylase (AH) (16%), ethylmorphone N-demethylase (EMND) (26%), and aminopyrine N-demethylase (AMND) (18%) activities and cytochrome P-450 (12%) and cytochrome b5 (10%) levels. Cd did not alter the hepatic microsomal heme level. Cd, however, significantly increased the hepatic microsomal p-nitroanisole O-demethylase (p-NAOD) (53%) activity. When Ni (59.5 mg NiCl2 . 6H2O/kg, sc) was administered to the guinea pigs 16 h prior to sacrifice, the metal significantly depressed AH (49%), p-NAOD (66%), EMND (47%), and AMND (37%) activities, and cytochrome P-450 (15%), cytochrome b5 (24%), and microsomal heme (28%) levels. For the combined treatment, animals received the single dose of Ni 56 h after the single dose of Cd and then were killed 16 h later. In these animals, significant inhibitions were noted in AH (51%), EMND (47%), and AMND (30%) activities, and cytochrome P-450 (15%), cytochrome b5 (26%), and microsomal heme (30%) compared to those of controls. In the case of p-NAOD activity, the influence was in favor of Ni, i.e., the inhibition was about 61% by the combined treatment. These results reveal that: 1. The response of all substrates of hepatic monooxygenases to Cd are not the same, possibly indicating differential regulation of cytochrome P-450 isozymes by Cd; 2. The inhibitory effect of Ni on hepatic monooxygenases is more profound than that of Cd; and 3. The combination of Cd and Ni does not have a synergistic effect of hepatic monooxygenases of the guinea pig

Çevredeki Alkifenoller ve Canlı Organizmalara Olan Zararlı Etkileri

The compounds of alkylphenolpolyethoxylates (APEs), one of the ubiquitous estrogenic environmental endocrine disrupters, are widely used as non-ionic surfactants or antioxidants in detergents, pesticides, herbicides, emulsifiers, paints, cosmetics, plasticwares and even in jet fuel. It has been reported that APEs and their derivatives called alkylphenols (APs) exert adverse effects on both aquatic and terrestrial organism. Moreover, they have been shown to have toxic, estrogenic and carcinogenic effects. Therefore, the occurrence of APEs and their derivatives in the environment as well as their structures, biodegradation, bioaccumulation, metabolic fate and their adverse effects on living organisms including human were reviewed from a complex literature to understand their potential danger to human being. From this review of literature, it is concluded that the pollution level of APEs or their derivatives in the environment are not lethal to the most organisms. However, the sub-lethal levels of these substances exert estrogenic and carcinogenic effects in almost all organisms including human being.Alkilfenol bileşikleri (AFEO) iyonic olmayan yüzey aktif maddesi olarak deterjanlarda, ot ve böcek ilaçlarında, kozmeiklerde, plastik eşyalarda emülsifikatörlerde, boyalarda ve hatta uçak yakıtlarında çok yaygın kullanılan östrojenik endokrin sistem bozuculardır. Alkilfenol adı verilen AFEO bilişiklerinin türevlerinin hem suda hem de karada yaşayan canlılara zararlı etkileri olduğu rapor edilmiştir. Ayrıca, bunların hem östrojenik, hem toksik hem de karinojenik etkileri olduğu ortaya konmuştur. Bu nedenle, AFEO bileşikleriri ve türevlerinin doğadaki varoluşları, yapıları, biyolojik bozunumları, biyolojik birikimleri, metbolize edilme yolları ve insan dahil bütün canlı organizmalara olan zararlı etkileri karmaşık literatür taranarak potnsiyel zararlı etkileri anlaşılmaya çalışılmıştır. Bu literatür taraması ile doğada bulunan AFEO bileşikleri ve türevlerinin miktarlarının öldürücü dozlarda olmadığı anlaşılmıştır. Fakat, bu bileşiklerin ve türevlerinin öldürücü olmayan dozlarının da insan dahil bütün canlı organizmalarda hem östrojenik hem de karsinojenik etkilerinin olduğu ortaya konmuştur

CHARACTERIZATION OF GLUTATHIONE S-TRANSFERASES FROM NEEDLES OF Pinus brutia Ten. TREES

Glutathione S-transferases (GST, E.C. 2.5.1.18) are generally dimeric and multifunctional enzyme family which catalyse the nucleophilic attack of the glutathione on lipophilic compounds with electrophilic centres. Since the 70's GSTs in plant species have been intensively studied, as their role discovered in herbicide detoxification. However, there is only a limited number of studies considering the GST enzyme composition from forest trees, especially not in Pinus brutia, Ten. The trees that exhibited healthy appearance were selected and all belong to the same altitude profile which is located in METU / Yalincak area (Ankara, Turkey). GST activities in the supernatant fractions prepared from needles of P.brutia were determined spectrophotometrically by using 1-chloro-2,4-dinitrobenzene, 2,3-dichloro-4-(2-methylene butyryl)-phenoxy acetic acid (ethacrynic acid), 1,2-dichloro-4-nitrobenzene, 1,2-epoxy-3-(p-nitrophenoxy) propane and p-nitrobenzyl chloride as substrates. Only 1-chloro-2,4-dinitrobenzene (160 +/- 10 nmoles min(-1) mg(-1)) and 1,2-dichloro-4-nitrobenzene (2.30 +/- 0.38 nmoles min(-1) mg(-1)) activities were detected and the rest were found as negligible. Accordingly, during purification of GSTs from needles of P.brutia, 1-chloro-2,4-dinitrobenzene was used as the substrate. Purification of GSTs was performed by sequential application of supernatant to gel filtration column chromatography on Sephadex G-25, anion exchange diethylaminoethyl cellulose column chromatography and S-hexylglutathione agarose affinity chromatography. After the final step of purification procedure, 1-chloro-2,4-dinitrobenzene conjugating activity of P. brutia cytosolic GSTs was purified about 15.45 fold with 1.95% yield. Sodium dodecyl sulfate polyacrylamide gel electophoresis results showed that the purified GST isozyme had an Mr of 24 kDa. With this study, we report for the first time the GST isozymes in a gymnosperm, P. brutia