Enhanced purification protocol for the angiotensin-converting enzyme from bovine systems and investigation of the in vitro effect of some active substances.

Angiotensin-converting enzyme (ACE, EC 3.4.15.1) synthesized by endothelial cells and responsible for the regulation of blood pressure was purified from the bovine lung with affinity chromatography method. The purification rate of the ACE of the bovine lung was calculated as 1748- fold. Optimum pH and optimum temperature for the purified ACE were found to be 7.6 and 35-40 °C, respectively. The purity and molecular weight of the ACE were designated with SDS-PAGE. The ACE was found to have three subunits with molecular weights of 57 kDa, 66 kDa, and 190 kDa. Then, the total molecular weight of the ACE was designated as 303 kDa with gel filtration chromatography. The effects of ACE inhibitors captopril, fosinopril, lisinopril, and beta-blockers propranolol, atenolol, and diuretic triamterene on ACE activity were studied. ACE inhibitors lisinopril, captopril, fosinopril, and diuretic triamterene demonstrated an inhibition effect on ACE activity. Beta-blockers indicated no effect on ACE. IC50 values of captopril, fosinopril, lisinopril, and triamterene from the graphical equation were calculated as 0.835 nM, 1.159 µM, 4.085 nM, and 227 µM, respectively. The inhibition type and Ki values of these compounds were determined from Lineweaver-Burk plots. Captopril, fosinopril, lisinopril, and triamterene demonstrated a non-competitive inhibition effect on ACE activity. Ki constants were found as 1.057 nM, 1.675 µM, 6.449 nM, and 419.5 µM, respectively. Captopril indicated the highest inhibitor effect with an IC50 value of 0.835 nM.

A novel nonenzymatic hydrogen peroxide amperometric sensor based on Pd@CeO2-NH2 nanocomposites modified glassy carbon electrode.

Herein, (3-aminopropyl)triethoxysilane functionalized cerium (IV) oxide (CeO2-NH2) supported Pd nanoparticles were synthesized. The nanocomposites were characterized using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and High-resolution transmission electron microscopy (HRTEM). The Pd@CeO2-NH2 showed better electrocatalytic response to the reduction of H2O2 than CeO2-NH2. The fabricated sensor exhibited two linear responses to the reduction of H2O2. The first one was from 0.001 to 3.276 mM with 0.47 µM of a limit of detection (LOD) (S/N = 3) and excellent sensitivity of 440.72 µA mM-1 cm-2 and the second one was from 3.276 to 17.500 mM with the sensitivity of 852.65 µA mM-1 cm-2 in the optimum conductions. Also, the sensor exhibited 91% of electrocatalytic activity toward H2O2 after having been used for 30 days and the reproducibility was also satisfactory. The sensor response to H2O2 was not affected by ascorbic acid, fructose, glycine, dopamine, arginine, mannose, glucose, uric acid, Mg+2, Ca+2, and phenylalanine at the studied potential. Also, the fabricated sensor was used to determine H2O2 in milk samples. The results show that the constructed sensor can be a promising devise for the determination of H2O2 in real samples.

Total phenolic content, ferric reducing and DPPH scavenging activity of Arum dioscoridis.

Arum dioscoridis, locally called 'Gavur pancari', is a wild plant the leaves of which have been used as vegetable and for preparing special soup which has a sour taste. This study was set up to determine in vitro antioxidant activities and total phenolic and flavonoid contents of different extracts of A. dioscoridis. Free radical scavenging activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing activity with different concentrations of ethanol, methanol, acetone and water extracts of the plant leaves. Total phenolic and flavonoid contents were widely variable depending on solvents. Ethanol and methanol extractions of the plant material showed better performances with respect to both phenolic and flavonoid contents, respectively. The highest phenolic and flavonoid contents of ethanol and methanol extracts were 100.890 mg/g GAE and 72.643 mg/g QE, respectively. The lower DPPH scavenging and ferric reducing activities were determined in comparison with previous reports and standard synthetic chemicals.