Murat Cankaya

The Inhibitory Effects of L-Adrenaline on Lactoperoxidase Enzyme Purified from Bovine Milk

L-Adrenaline belongs to a group of compounds known as catecholamines, which play an important role in the regulation of the physiological process in living organisms. In the present study, the inhibitory effect of L-adrenaline on lactoperoxidase was examined. Lactoperoxidase (E.C.1.11.1.7) was purified from bovine milk with three consecutive steps: Amberlite CG-50 resin, CM-Sephadex C-50 ion-exchange, and Sephadex G-100 gel filtration chromatography. Lactoperoxidase was purified with a yield of 42.18%, a specific activity of 30.33 EU/mg proteins, and 20.77 purification fold. Enzyme purity was determined with SDS-PAGE, where a single band was observed. The Rz (A412/A280) value for lactoperoxidase was 0.9. The effect of L-adrenaline on lactoperoxidase was determined using ABTS as a chromogenic substrate. The half maximal inhibitory concentration (IC50) value and an inhibition constant (Ki) values for L-adrenaline were 34.5 and 2.26 μM, respectively. L-Adrenaline was found to be a non-competitive inhibitor.

Inhibition profile of a series of phenolic acids on bovine lactoperoxidase enzyme

Abstract Lactoperoxidase (LPO) catalyzes the oxidation of numerous of organic and inorganic substrates by hydrogen peroxide. It has very vital activity in the innate immune system by decreasing or stopping the activation of the bacteria in milk and mucosal secretions. This study’s purpose was to investigate in vitro effect of some phenolic acids (ellagic, gallic, ferulic, caffeic, quercetin, p-coumaric, syringic, catechol and epicatechin) on the purified LPO. This enzyme was purified from milk by using different methods such as Amberlite CG-50 resin, CM-Sephadex C-50 ion-exchange and Sephadex G-100 gel filtration chromatography. LPO was purified 28.7-fold with a yield of 20.03%. We found phenolic acids have inhibition effects on bovine LPO enzyme to different concentrations. Our study showed lower concentrations of caffeic acid, ferulic acid and quercetin exhibited much higher inhibitory effect on enzyme, so these three of them were clearly a more potent inhibitor than the others were. All of compounds were non-competitive inhibitors.

Four diclofenac complexes with cobalt(II) and nickel(II) ions: synthesis, spectroscopic properties, thermal decompositions, crystal structures, and carbonic anhydrase activities

Cobalt(II) and nickel(II) complexes with diclofenac (dicl) in the presence of nitrogen-donor 3-picoline (3-pic) and 1-(2-aminoethyl)pyrrolidine (2-aepyr), [Co(dicl)2(3-pic)2] (1), [Ni(dicl)2(3-pic)2(H2O)2] (2), [Co(dicl)2(2-aepyr)2] (3), and [Ni(dicl)2(2-aepyr)2] (4), have been synthesized and characterized by FT-IR, UV–Vis, elemental and thermal analysis. The crystal structures of 1 and 3 have been determined by single-crystal X-ray diffraction; phase purity of complexes has been proved by powder X-ray diffraction analysis. Structural analyses have demonstrated that 1 and 3 are mononuclear and Co(II) ions have distorted octahedral environments. In 1, dicl is bidentate, whereas in 2, 3, and 4 dicl is monodentate. Dicl ligands are coordinated to metal(II) ions with O of carboxylate. Therefore, IR spectra of all complexes display {ν(OCO)asym and ν(OCO)sym} of dicl. The calculated Δν(OCO) values are consistent with the presence of monodentate (>200) and bidentate (<200) carboxylate. Compounds 2, 3, and 4 exhibit inhibition effects on human carbonic anhydrase-I. Compounds 3 and 4 show high thermal stability compared with 1 and 2. Graphical Abstract

Effects of some drugs on human cord blood erythrocyte carbonic anhydrases I and II: an in vitro study

In the present study, we purified hcbCA I and II from human cord blood erythrocytes using by Sepharose-4B-l tyrosine-sulfanilamide affinity gel chromatography. Also, it was checked the inhibition effects of ampicillin sulfate, ceftriaxone, ceftizoxime and ranitidine on hcbCA I and hcbCA II. IC50 values for ceftriaxone, ceftizoxime and ranitidine were found to be 27.l, 79.4 and 55.5 µM for hcbCA I and of 21.0, 79.1 and 66.1 µM for hcbCA II, respectively. According to these results, Ampicillin sulfate inhibited only hcbCAII and IC50 values of this antibiotic was found to be 56.8 µM. All these substances were found non-competitive inhibitors. It is important to study the inhibition effects of these drugs on hcbCA I and II izoenzymes. Because, pregnant woman is take all of these substance. For this reason, these drugs should be carefully used and the dosage should be very well ordered to minimize side effects.

Interaction between axons and specific populations of surrounding cells is indispensable for collateral formation in the mammillary system.

Background An essential phenomenon during brain development is the extension of long collateral branches by axons. How the local cellular environment contributes to the initial sprouting of these branches in specific points of an axonal shaft remains unclear. Methodology/Principal Findings The principal mammillary tract (pm) is a landmark axonal bundle connecting ventral diencephalon to brainstem (through the mammillotegmental tract, mtg). Late in development, the axons of the principal mammillary tract sprout collateral branches at a very specific point forming a large bundle whose target is the thalamus. Inspection of this model showed a number of distinct, identified cell populations originated in the dorsal and the ventral diencephalon and migrating during development to arrange themselves into several discrete groups around the branching point. Further analysis of this system in several mouse lines carrying mutant alleles of genes expressed in defined subpopulations (including Pax6, Foxb1, Lrp6 and Gbx2) together with the use of an unambiguous genetic marker of mammillary axons revealed: 1) a specific group of Pax6-expressing cells in close apposition with the prospective branching point is indispensable to elicit axonal branching in this system; and 2) cooperation of transcription factors Foxb1 and Pax6 to differentially regulate navigation and fasciculation of distinct branches of the principal mammillary tract. Conclusions/Significance Our results define for the first time a model system where interaction of the axonal shaft with a specific group of surrounding cells is essential to promote branching. Additionally, we provide insight on the cooperative transcriptional regulation necessary to promote and organize an intricate axonal tree.

2-Amino-3-cyanopyridine derivatives as carbonic anhydrase inhibitors.

Carbonic anhydrases (CAs, EC 4.2.1.1) are ubiquitous enzymes that catalyze the hydration of CO2 to bicarbonate and protons. Inhibition of CAs has been clinically exploited for the treatment of various classes of diseases for decades, but investigating new classes of inhibitors continues to be important. We have synthesized a series of 2-amino-3-cyano-4-heteroaryl (5a–l) compounds and characterized the structures by NMR, IR and elemental analyses. We tested the ability of these compounds to inhibit two metalloenzyme human carbonic anhydrase (hCA, EC 4.2.1.1) isozymes, hCA I and hCA II. Compounds 5d and 5b showed the best inhibition activity against hCA I (IC50: 33 and 34 µM, respectively), and compound 5d showed the best activity against hCA II (IC50: 56 µM).

Oxidative and antioxidative status in the endometrium of patients with benign gynecological disorders

AIM Oxidative stress and impaired antioxidative system are implicated in the development of many disease states including gynecological diseases. In the present study, we aimed to investigate the oxidative-antioxidative status in endometrium of patients diagnosed with benign gynecological disorders. METHODS Samples of endometria and blood were obtained from 65 patients admitted to our center for abnormal uterine bleeding or postmenopausal bleeding. Endometrial biopsy was performed for the evaluation of histopathology and oxidative-antioxidative status in endometrial tissue. Based on histological examination, subjects were divided into groups as follows: normal controls (n=15); patients with endometrial polyps (n=20); patients with uterine myoma (n=10) patients with chronic endometritis (n=10), and patients with atrophic endometrium (n=10). Activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and total antioxidant status (TOS), total oxidant status (TAS) were assessed. RESULTS Compared to the normal controls, nonsignificant changes (decrease or increase) were detected in antioxidant enzyme activities, TAS and TOS in the examined groups. Additionally, between TAS and TOS, we also found a strong positive correlation in normal and chronic endomethritis groups and a moderate positive correlation uterine myoma, endometrial polyps and endometrial atrophy groups. CONCLUSION Even though, our results do not allow to conclude how oxidative and antioxidative status are influenced in benign gynecological disorders, these findings may provide a basis for the further researches.

Prevention of infertility induced by ovarian ischemia reperfusion injury by benidipine in rats: Biochemical, gene expression, histopathological and immunohistochemical evaluation

INTRODUCTION Benidipine has been reported to prevent the ischemia/reperfusion (I/R) damage in heart tissue and to suppress oxidant and proinflammatory cytokine production, increased by I/R. However, There was no information about the effects of benidipine on I/R injury in the ovary and the damage of I/R-induced infertility. OBJECTIVES The aim of the study was to investigate the effects of benidipine on bilateral ovarian I/R injury and whether or not effective in the treatment of I/R-induced infertility in rats. METHOD Forty-eight females, albino Wistar rats were randomly divided into 4 groups: IRC group (ovarian I/R group, n=12), IRB-2 group (ovarian I/R+2mg/kg benidipine group, n=12), IRB-4 group (ovarian I/R+4mg/kg benidipine group, n=12) and HG group (healthy group with sham operation, n=12). In IRB-2 and IRB-4 groups, two hours ischemia and two hours reperfusion was performed following orally benidipine administration. After this I/R procedure, 6 rats from each group performed bilateral overectomy. Ovarian levels of malondialdehyde (MDA) and total glutathione (tGSH), ovarian gene expressions of interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) and also apoptosis were evaluated. The other 6 rats from each group were put in together with six male rats in separated cages for 2 months in order to reproduce. During this period, rats which did not become pregnant were accepted as infertile. RESULTS MDA levels, expressions of TNF-α and IL-1β in IRC group were significantly higher than in the SGA group and tGSH was decreased. In total, 4mg/kg benidipine has better prevented ovaries from the increase of oxidants and proinflammatory cytokines, the decrease of antioxidants than 2mg/kg benedipine. In the histopathological examination hemorrhage, congestion, follicle degeneration, neutrophil infiltration and necrosis were seen in ovarian tissue of IRC group. Only dilated and congested blood vessels were found in the IRB-2 group. No histopathological finding was encountered in the IRB-4 group. I/R caused infertility in rats. In total, 4mg/kg benidipine prevented from infertility better than the dose of 2mg/kg benedipine. CONCLUSION In total, 4mg/kg benidipine reduced I/R injury and I/R-related infertility more significantly compared to 2mg/kg benedipine in rat ovaries.

Synthesis of 2‐amino‐3‐cyanopyridine derivatives and investigation of their carbonic anhydrase inhibition effects

The conversion of carbon dioxide (CO2) and bicarbonate (HCO3−) to each other is very important for living metabolism. Carbonic anhydrase (CA, E.C.4.2.1.1), a metalloenzyme familly, catalyzes the interconversion of these ions (CO2 and HCO3−) and are very common in living organisms. In this study, a series of novel 2‐amino‐3‐cyanopyridines supported with some functional groups was synthesized and tested as potential inhibition effects against both cytosolic human CA I and II isoenzymes (hCA I and II) using by Sepharose‐4B‐l‐tyrosine‐sulfanilamide affinity chromatography. The structural elucidations of novel 2‐amino‐3‐cyanopyridines were achieved by NMR, IR, and elemental analyses. Ki values of the novel synthesized compounds were found in range of 2.84–112.44 μM against hCA I and 2.56–31.17 μM against hCA II isoenzyme. While compound 7d showed the best inhibition activity against hCA I (Ki: 2.84 μM), the compound 7b demonstrated the best inhibition profile against hCA II isoenzyme (Ki: 2.56 μM).

The preventive role of levosimendan against bleomycin-induced pulmonary fibrosis in rats

BACKGROUND In this study, the effects of levosimendan used in the treatment of acute congestive heart failure upon pulmonary fibrosis in rats induced with bleomycin (BL) were analyzed. METHODS A total of 33 male Sprague-Dawley type rats were categorized into five groups randomly. About 2.5U/kg BL was intratracheally administered to the rats in the BL, BL+L1, BL+L2, and BL+L3 groups, and 0.9% saline was intratracheally administered at the same rate to the control group. 0.3, 1, and 3mg/kg levosimendan was intraperitoneally administered to the BL+L1, BL+L2, and BL+L3 groups, respectively. Blood and tissue samples were taken from the rats euthanized to determine the changes in erythrocyte enzyme activities and to conduct histopathological evaluations after 14 days. With values between 0 and 3, histopathological scoring damage was assessed by the presence of inflammation and fibrosis in a semiquantitative manner. RESULTS Compared with those in the C group, glutathione reductase (GR) and Catalase (CAT) enzymes decreased in the BL group; compared with that in the BL group, GR increased in the BL+L1 and BL+L3 groups, 6-phosphogluconate dehydrogenase (6PGD) increased in the BL+L3 group, and CAT increased in the BL+L2 and BL+L3 groups (p<0.05). In the histopathological evaluation, fibrosis occurred in all rats in the BL group, and tissue damage was noticed to be generally less in the BL+L1, BL+L2, and BL+L3 groups (p<0.001). CONCLUSIONS The results obtained from biochemical and histopathological evaluations indicate that levosimendan had an anti-fibrotic effect without a dose-dependent response on pulmonary fibrosis.