Mete Özkurt

Indomethacin based new triazolothiadiazine derivatives: Synthesis, evaluation of their anticancer effects on T98 human glioma cell line related to COX-2 inhibition and docking studies.

In the current work, new 1,2,4-triazolo[3,4-b]-1,3,4-thiadiazine derivatives (1-8) were synthesized via the ring closure reaction of 2-bromoacetophenone derivatives with 4-amino-5-[(5-methoxy-2-methyl-1-(4-chlorobenzoyl)-1H-indol-3-yl)methyl]-2,4-dihydro-3H-1,2,4-triazol-3-thione, which was obtained via the solvent-free reaction of indomethacin with thiocarbohydrazide. MTT assay was carried out to determine the cytotoxic effects of the compounds on T98 human glioma cell line. Among these compounds, 3-[5-methoxy-2-methyl-1-(4-chlorobenzoyl)-1H-indole-3-yl)methyl]-6-(4-methylphenyl)-7H-1,2,4-triazolo[3,4-b]-1,3,4-thiadiazine (8) was found to be the most effective compound and therefore flow cytometric method was performed to investigate the apoptotic effect of compound 8. The apoptosis stimulating percentages of compound 8 in comparison with the control group at 50 and 100 μM doses were calculated as 11% and 12%, respectively. Besides, real-time PCR assay was carried out to determine the effects of compound 8 on COX-2, caspase 3, 8 and 9, cytochrome c mRNA levels. According to the real-time PCR analysis, compound 8 reduced COX-2 mRNA levels significantly when compared with the control group, whereas the compound did not cause any significant change in other parameters (Caspase 3, 8, 9, cytochrome c). The docking study suggested that the COX-2 inhibitory effects of compound 8 and indomethacin were similar in the catalytic active site of COX-2. These results indicated that compound 8 showed dose-dependent anticancer activity via the inhibition of COX-2 pathway.

Association of angiotensin converting enzyme (ACE) gene I/D polymorphism and polycystic ovary syndrome (PCOS).

This study was conducted in Turkish patients with polycystic ovary syndrome to determine the frequency of I/D polymorphism genotypes of angiotensin converting enzyme gene, and to examine the role of this polymorphism in polycystic ovary syndrome development. Genomic DNA obtained from 200 persons (100 patients with polycystic ovary syndrome and 100 healthy controls) was used in the study. DNA was multiplied by polymerase chain reaction using I and D allele-specific primers. Polymerase chain reaction products were assessed with a charge coupled device (CCD) camera by being exposed to 2% agarose gel electrophoresis. There was statistically significant difference between the groups with respect to genotype distribution (p<0.001). The D allele frequency was indicated as 68% and I allele was as 32% in the patients, whereas it was 51.5-48.5% respectively in the control group. As a result of our study we may assert that angiotensin converting enzyme gene I/D polymorphism DD genotype should be considered as a genetic marker in polycystic ovary syndrome development in this Turkish study population.

Effect of αlipoic acid and silymarin on bladder outlet obstruction

The aim of the present study was to determine whether the treatment of obstructed rat bladders with αlipoic acid (ALA) and silymarin reverses the biochemical and physiological responses to bladder outlet obstruction (BOO). A total of 32 adult Sprague Dawley rats were divided into four groups (n=8 per group): sham (placebo surgery) animals with no treatment (group 1); control animals with surgically induced BOO (group 2); obstructed rats treated with ALA (group 3); and obstructed rats treated with silymarin (group 4). Histological evaluation, bladder weights, collagen structure, TdT-mediated biotin nick end-labeling (TUNEL), inducible nitric oxide sentase (iNOS) mRNA levels, malondialdehyde (MDA) levels and tumor necrosis factor (TNF) levels were investigated. The ALA-treated group had similar bladder weights, collagen levels and TUNEL positivity and decreased iNOS levels compared with the control group, while the silymarin group exhibited further differences. Serum MDA and TNF-α levels were both decreased in the ALA and silymarin groups. ALA treatment reduced the increased oxidative stress and bladder inflammation caused by BOO and may contribute to the protection of bladder function.

Plasminogen activator inhibitor type-1 gene 4G/5G polymorphism and polycystic ovary syndrome.

AIM This study was conducted in Turkish patients with polycystic ovary syndrome (PCOS) to determine the frequency of 4G/5G polymorphism genotypes of plasminogen activator inhibitor type-1 gene and to examine the role of this polymorphism in PCOS development. MATERIALS AND METHODS Genomic DNA obtained from 200 persons (100 patients with PCOS and 100 healthy controls) was used in the study. DNA was amplified by polymerase chain reaction using 4G and 5G allele-specific primers. Polymerase chain reaction products were assessed with charged-couple device camera after exposing to 2% agarose gel electrophoresis. RESULTS No statistically significant difference between the groups with respect to genotype distribution was found (p>0.05) in the study. The 4G and 5G allele frequencies were indicated as 51.5% and 48.5% in patients, respectively, whereas this was 51%-49% in the control group. CONCLUSION Consequently, it has been established by this study that the 4G/5G polymorphism genotypes of plasminogen activator inhibitor type-1 gene do not play a role in the development of PCOS in the Turkish population.

Erythropoietin Protects the Kidney by Regulating the Effect of TNF-α in L-NAME-Induced Hypertensive Rats

Background/Aims: Hypertension is the leading cause of death worldwide. Chronic high blood pressure induces inflammation. Tumor necrosis factor (TNF)-α plays a major role in inflammation and also depresses the synthesis of erythropoietin, which exerts protective effects on tissue; however, the mechanism is still unclear. We investigated the protective effect of erythropoietin against tissue damage caused by hypertension in the kidney and whether this effect was suppressed by TNF-α. Methods: First, we detected the optimum chronic dose for darbepoetin-α (Depo), which is a long-acting erythropoietin analog for rats. We separated 60 female adult rats into 6 groups: control, Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME), L-NAME+Depo, L-NAME+Remicade (an anti-TNF-α antibody), L-NAME+Depo+Remicade, Depo, and control. After 1 month of treatment, we measured cardiovascular parameters, took blood samples, sacrificed the rats, and removed kidneys for analyses. Results: The apoptotic index and the plasma and kidney mRNA levels of TNF-α increased in the L-NAME group and decreased in all other treatment groups. Macrophage accumulation increased in the L-NAME and L-NAME+Remicade groups, while it decreased in the Depo group. The mRNA abundance of TNF receptor 1 (TNFR1) decreased slightly in the Depo group and TNFR2 increased significantly in the same group. Conclusion: Erythropoietin protects kidney tissue against hypertension by preventing the apoptotic effects of TNF-α by blocking macrophage accumulation, decreasing TNF-α levels, and switching the TNF-α receptors from the apoptotic receptor TNFR1 to the proliferative receptor TNFR2.

Vasoactive intestinal peptide, a promising agent for myopia?

AIM To investigate the role of vasoactive intestinal peptide (VIP) in form-deprivation myopia (FDM). METHODS FDM was created in three groups of eight chicks by placing a translucent diffuser on their right eyes. Intravitreal injections of saline and VIP were applied once a day into the occluded eyes of groups 2 and 3, respectively. Retinoscopy and axial length (AL) measurements were performed on the first and 8th days of diffuser wear. The retina mRNA levels of the VIP receptors and the ZENK protein in right eyes of the three groups and left eyes of the first group on day 8 were determined using real time polymerase chain reaction (PCR). RESULTS The median final refraction (D) in right eyes were -13.75 (-16.00, -12.00), -11.50 (-12.50, -7.50), and -1.50 (-4.75, -0.75) in groups 1, 2, and 3, respectively (P<0.001). The median AL (mm) in right eyes were 10.65 (10.00, 11.10), 9.90 (9.70, 10.00), and 9.20 (9.15, 9.25) in groups 1, 2, and 3, respectively (P<0.001). The median delta-delta cycle threshold (CT) values for the VIP2 receptors were 1.07 (0.82, 1.43), 1.22 (0.98, 1.65), 0.29 (0.22, 0.45) in right eyes of groups 1, 2, and 3, and 1.18 (0.90, 1.37) in left eyes of group 1, respectively (P=0.001). The median delta-delta CT values for the ZENK protein were 1.07 (0.63, 5.03), 3.55 (2.20, 5.55), undetectable in right eyes of groups 1, 2, and 3 and 1.89 (0.21, 4.73) in left eyes of group 1, respectively (P=0.001). CONCLUSION VIP has potential inhibitory effects in the development of FDM.