İnsan eritrosit karbonik anhidraz izoenzimleri üzerine bazı kardiyovasküler terapötiklerin inhibisyon etkilerinin incelenmesi

Carbonic anhydrase enzyme plays a vital role in metabolic events such as acid-base regulation and respiration. In our research, it is tried to determine the inhibitory influences of the cardiovascular therapeutics esmolol hydrochloride, amiodarone hydrochloride and lidocaine hydrochloride on human erythrocytes carbonic anhydrases (hCA I and II). In accordance with this purpose, carbonic anhydrase isoenzymes were purified from human erythrocytes by using affinity chromatography method. Enzyme purity was checked by SDS-PAGE electrophoresis method. After the carbonic anhydrase enzymes were purified, the inhibitory affects of cardiovascular therapeutics on these enzymes, using esterase activity, which is the method of measuring in vitro activity, were examined. The three cardiovascular therapeutics dose-dependently decreased activity of hCAs. IC50 values of amiodarone hydrochloride, esmolol hydrochloride and lidocaine hydrochloride were found to be, respectively, 0.91 mM, 5 mM, 5.8 mM for hCA-I and 0.41 mM, 3.5 mM, and 6.36 mM for hCA-II. Our results proved that, under in vitro conditions, cardiovascular therapeutics significantly inhibit human CA-I and II activities. So, irregular use of these medicines may cause serious adverse effects in terms of human health.Karbonik anhidraz enzimi, asit-baz düzenlemesi ve solunum gibi metabolik olaylarda hayati rol oynar. Bu çalışmada, bazı kardiyovasküler terapötiklerin (esmolol hidroklorür, amiodaron hidroklorür, lidokain hidroklorür) insan eritrositleri karbonik anhidraz I ve II izoenzimleri (hCA-I ve hCA-II) aktiviteleri üzerine inhibisyon etkilerinin belirlenmesi amaçlanmıştır. Bu amaçla, insan eritrositlerinden afinite kromatografisi yardımıyla karbonik anhidraz izoenzimleri (CA-I ve CA-II) saflaştırıldı. Enzim saflıkları SDS-PAGE ile kontrol edildi. Saflaştırmadan sonra, kardiyovasküler terapötiklerin hCA-I ve hCA-II aktivitesi üzerindeki inhibitör etkileri, in vitro olarak esteraz yöntemi kullanılarak farklı ilaç konsantrasyonlarında belirlendi. Üç kardiyovasküler terapötik doza bağımlı olarak enzim aktivitelerini azalttı. Amiodaron hidroklorür, esmolol hidroklorür ve lidokain hidroklorür’ün IC50 değerleri hCA-I için sırasıyla 0.91 mM, 5 mM, 5.8 mM olarak, hCA-II için sırasıyla 0.41 mM, 3.5 mM, 6.36 mM olarak bulundu. Bulgularımız, kardiyovasküler terapötiklerin in vitro koşullarda hCA-I ve hCAII aktivitesini önemli ölçüde inhibe ettiğini göstermektedir. Bu nedenle, bu ilaçların kontrolsüz kullanımı insan sağlığı için ciddi yan etkilere neden olabilir

Bazı metallerin Minekop (Umbrina Cirrosa) solungacı karbonik anhidraz aktivitesi üzerine etkisi

Metal toxicity causes oxidative stress in fish. This situation is a potential risk factor for humans and other living feeding on contaminated fish. In this study, the inhibition effects of heavy metals on carbonic anhy- drase enzyme from the corb fish gill were investigated. The carbonic anhydrase enzyme was purified from gill of corb fish with a specific activity of 2093,9 EUmg-1 and 86,51% yield and approximately 160 fold using Sep- harose 4B–L-tyrosine sulfanilamide affinity chromatography method. SDS–polyacrylamide gel electrophoresis showed a single band corresponding to a molecular weight of approximately 30,8 kDa. Inhibitory effects of metals (Ag+, Cu2+, Pb2+, Zn2+, Cd2+, Ni2+) on CA activity were determined at different concentrations using the hydratase method under in vitro conditions. Consequently, in vitro inhibition rank order was determined as Ag+> Cu2+> Pb2+> Zn2+ > Cd2+> Ni2+. From these results, we showed that Ag+ is the most potent inhibitor of CA enzyme

Influence of pesticide exposure on carbonic anhydrase II from sheep stomach

Carbonic anhydrase (CA) is a widely distributed enzyme and has a crucial role in the cells, tissues and organs of living organisms. It is found that CA-II is one of the most abundant CA isoenzymes in the gastrointestinal system. It plays an important role in the gastric acid secretion in stomach. In this study, we purified CA-II isoenzyme from sheep stomach with a 615.2 purification fold, 78% purification yield and 5562.02 specific activity. Moreover, the in vitro effects of some commonly used pesticides including chlorpyrifos, cypermethrin, dichlorvos, glyphosate isopropylamine and lambda cyhalomethrin on the enzyme activity were investigated. Of these compounds, glyphosate isopropylamine and dichlorvos showed an inhibition on CA-II esterase activity. They have IC50 values of 0.155 mM and 2.690 mM and Ki values of 0.329 mM and 3.654 mM, respectively. Both glyphosate isopropylamine and dichlorvos inhibited CA-II isoenzyme in a noncompetitive manner

Effects of subclinical mastitis on milk somatic cells and milk vitamin C levels in cows

Çalışma, subklinik mastitisli ineklerde süt ve süt hücrelerinde Vitamin C düzeylerinin belirlenmesi amacıyla yapılmıştır. Süt örneklerinde somatik hücre sayımı yapıldı ve örnekler kontrol (1-87 x 1000 hücre), mastitli 1. grup (154-380 x 1000 hücre), 2. grup (418-812 x 1000 hücre), 3. grup (914-1928 x 1000 hücre) ve 4. grup (2614-8050 x 1000 hücre) olacak şekilde gruplandırıldı (n=12). Süt hücrelerinde ve süt serumunda (yağı ve hücreleri alınmış) Vitamin C düzeyleri belirlendi ve Vitamin C ve süt somatik hücre sayısı arasındaki korelasyonlar hesaplandı. Ayrıca alınan süt numunelerinden mikrobiyolojik ekim yapılarak, etken izolasyonu ve identifikasyonu yapıldı. Subklinik mastitisli sütlerde (1, 2, 3, ve 4. gruplar) µg/106 hücredeki Vitamin C düzeyleri kontrol grubundan düşük olarak bulunurken (p0,05) göre düşük olduğu görüldü. Ayrıca süt serumu Vitamin C düzeyleri ile süt somatik hücreleri arasında negatif bir korelasyon saptandı (r=-0,420, p<0,01 n=60). Sonuç olarak subklinik mastitisin derecesi ile ilgili olarak somatik hücre sayısının arttığı, süt serumu Vitamin C düzeylerinin düştüğü, birim hücre başına düşen Vitamin C düzeylerinin azaldığı, Vitamin C düzeyleri ile mastit arasında bir bağıntının olduğu belirlendi.In this study, it was aimed to determine the levels of Vit C in milk and milk cells of subclinical mastitic cows. Somatic cell count was determined in the quarter milk samples of all animals and the groups were scored control (1-87 x 1000 cells), mastitic 1st group (154-380 x 1000 cells), 2nd group (418-812 x 1000 cells), 3rd group (914-1928 x 1000 cells) and 5th group (2614-8050 x 1000 cells) (n=12). Milk serum and milk cell Vitamin C levels were determined and the correlation between Vitamin C levels and somatic cell count were evaluated. Also microbiological diagnosis was conducted. In subclinical mastitic milks, Vitamin C levels (&micro;g/106 cell) were lower compared to control group (p&lt;0,001) and positive correlation was determined between milk cell Vitamin C levels and somatic cell count (r=0,469, p&lt;0,001 n=60). As regards milk serum, Vitamin C levels were lower in the 5th group compared to the other mastitic (p&lt;0,05) and control groups (p&gt;0,05). There was a negative correlation between milk serum Vitamin C levels and somatic cell count (r=-0,420, p&lt;0,01 n=60). In conclusion, it was found that somatic cell count was elevated with the mastitis levels, gradually, Vitamin C levels per cell and in milk serum decreased in mastitis. It is also found that there was a correlation between Vitamin C levels and mastitis