Irfan Degirmenci

The association of MMP-9 enzyme activity, MMP-9 C1562T polymorphism, and MMP-2 and -9 and TIMP-1, -2, -3, and -4 gene expression in lung cancer.

AIM To investigate the association of gene expression of MMP-2 and -9, and TIMP-1, -2, -3, and -4 and polymorphism frequencies of MMP-9 C1562T and plasma MMP-9 enzyme activity in lung cancer patients. METHODS In this study, DNA and RNA samples were extracted from peripheral blood of 300 subjects (200 lung cancer patients and 100 controls). MMP-9 C1562T polymorphism was determined using restriction fragment length polymorphism (RFLP) method, and expression of MMP-2 and -9, TIMP-1, -2, -3, and -4 was determined using reverse transcriptase polymerase chain reaction. Plasma MMP-9 enzyme activity levels were measured using enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION The frequencies of C1562T genotypes were found to be CC 67%, CT 30%, and TT 3% in the control group and CC 75%, CT 24%, and TT 1% in the patient group. It was determined that CC genotype frequency increases significantly in patients according to control group. Plasma MMP-9 enzyme activity levels increased in patients with lung cancer compared to the control group. The cut-off value of MMP-9 enzyme activity was determined as 7.76 ng/mL by receiver operating characteristics curve analysis. The sensitivity, specificity, positive predictive value, and negative predictive value were 77%, 51%, 75.9%, and 52.6%, respectively. The expression of MMP-2 and TIMP-1 was found to be higher in lung cancer patients. Finally, we claim that determination of MMP-9 enzyme levels and expression of MMP-2 and -9 and TIMP-1 can be used as a marker in lung cancer.

Effects of various agents on DNA fragmentation and telomerase enzyme activities in adenocarcinoma cell lines

Natural compounds such as resveratrol, tannic acid, and quercetin may help to treat cancer. Tamoxifen is a non-steroidal anti-estrogen drug widely used in the treatment of patients with estrogen receptor-positive breast cancer. The aim of the study was to compare the effects of these natural compounds and tamoxifen in colon adenocarcinoma (CaCo-2) and breast adenocarcinoma (MCF-7) cell lines, on telomerase enzyme activity, cell viability, number of cells and DNA fragmentation. In this study to determine telomerase enzyme activity was used PCR–ELISA kit. To determine cell viability and number of cells were used tripan blue stain. DNA fragmentation was determined by DNA ladder isolation kit. Tannic acid was more effective than resveratrol, with respect to reduction in telomerase activity, cell viability and cell count in breast adenocarcinoma. Tannic acid and tamoxifen was more effective than resveratrol and quercetin telomerase activity, cell viability and cell count in colon adenocarcinoma. Flavonoids such as resveratrol, tannic acid and quercetin which was studied on, has benefical effects on cancer therapy. These effects such as decreasing telomerase enzyme activity, cell viability and number of cells and inducing DNA fragmentation (apoptosis) must be studied for assist to develop new therapeutic pathways. There should be much more sudies in order to discover resveratrol, tannic acid and quercetin and other potential medicines.

Investigation of Association Between Angiotensin-Converting Enzyme Gene Insertion/Deletion Polymorphism Frequency in Turkish Patients with Schizophrenia

AIM the insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme (ACE) gene is of increasing interest in etiology and treatment of various neuropsychiatric disorders. The present study aimed at detecting the incidence of ACE gene I/D polymorphism in patients with schizophrenia living in the Eskisehir region (Turkey) and also at determining whether this illness could be associated to ACE gene I/D polymorphism and serum ACE concentrations. METHODS in our study, genomic DNA was studied in a total of 237 individuals, 132 of them having been diagnosed as patients with schizophrenia and 105 of them being used as control subjects. In addition, sera from 31 patients with schizophrenia and 26 healthy subjects were used to compare serum ACE concentrations. By using polymerase chain reaction, we determined the frequency of ACE gene I/D polymorphism and measured the serum ACE concentrations by ELISA. RESULTS AND CONCLUSION distribution of ACE gene I/D polymorphism and allele frequencies between the control group genotype proportions (II 19%, ID 44%, DD 37%) and the patient group (II 19%, ID 45%, DD 36%) were not significantly different. Serum ACE concentrations were 293.15 ± 23.29 ng/mL in the control group and 362.61 ± 19.96 ng/mL in the patients. It was observed that serum ACE concentrations significantly increased in patients with schizophrenia compared with those of the control group (p < 0.05). However, no significant difference could be observed according to genotypes in serum ACE concentration.

Antioxidant effects of melatonin and coenzyme Q10 on oxidative damage caused by single-dose ochratoxin A in rat kidney.

In the study, the effects of relatively high single-dose of Ochratoxin A (OTA) and the antioxidant effects of Melatonin (Mel) and Coenzyme Q10 (CoQ10) on OTA-induced oxidative damages in rats were investigated. A total of 28 male Sprague-Dawley rats were divided into four groups of 7 rats each: Control, OTA, Mel+OTA and CoQ10+OTA groups. Malondialdehyde (MDA) levels in the plasma and glutathione (GSH) levels in whole blood were measured; kidneys (for histological inspection and for apoptosis detection by TUNEL method) and bone marrow samples (for chromosome aberration and mitotic index) were taken. The rats in the OTA group showed limited degeneration of tubular cells. In some tubules karyomegaly, desquamated cells and vacuolization were observed by light microscopy. Mel and CoQ10 treatment significantly reduced the severity of the lesions. MDA levels of the OTA group were significantly higher than the control, OTA+Mel and OTA+CoQ10 groups, while GSH levels were significantly lower than the control, OTA+Mel and OTA+CoQ10 groups. Higher incidences of apoptotic bodies were observed in the kidneys of the OTA group although OTA administration did not significantly change the incidence of apoptotic bodies when compared to the control and antioxidant administrated groups. Although the percentage of the mitotic index was lowest in the OTA group, no statistical difference was found among the groups. Additionally, OTA had no numerical and structural significant effects on chromosomes. It was observed that single-dose OTA administration caused oxidative damages in rat kidney and Mel or CoQ10 treatment appeared to ameliorate the OTA-induced tissue injuries.

Plasminogen Activator Inhibitor Type-1 Gene 4G/5G Polymorphism in Turkish Adult Patients with Asthma

AIM This study has been performed on asthmatic patients in the Turkish population to determine the frequency of 4G/5G polymorphism genotypes of plasminogen activator inhibitor type-1 gene, and with the aim of examining the role of this polymorphism in asthma development. METHODS Genomic DNA obtained from 165 persons (98 patients with asthma and 67 healthy controls) was used in the study. DNA was multiplied with polymerase chain reaction using 4G and 5G allele-specific primers. Polymerase chain reaction products were assessed with CCD camera by being exposed to 2% agarose gel electrophoresis. Results were evaluated with chi-square test. RESULTS No statistically significant difference between the groups with respect to genotype distribution was found (p > 0.05) in the study. The 4G allele frequency was indicated as 48% and 5G allele was as 52% in patients, whereas this was 50-50% in the control group. CONCLUSION It has been established by this study that 4G/5G polymorphism genotypes of plasminogen activator inhibitor type-1 gene do not play a role in the development of asthma in the Turkish population.

Investigation of Association between Plasminogen Activator Inhibitor Type-1 (PAI-1) Gene 4G/5G Polymorphism Frequency and Plasma PAI-1 Enzyme Activity in Patients with Acute Stroke

AIM This study was carried out to determine if there is any association between plasminogen activator inhibitor type-1 (PAI-1) gene 4G/5G polymorphism and plasma PAI-1 enzyme activity in acute stroke patients. METHODS In this study, 333 genomic DNAs (from 253 acute stroke patients and 80 healthy subjects) were analyzed. Genomic DNAs were prepared from peripheral blood using a saline method. These DNAs were amplified by PCR method using primers specific for 4G and 5G alleles. PCR products were separated by 2% agarose gel electrophoresis and visualized by a charge coupled device (CCD) camera. PAI-1 enzyme activities were measured by ELISA method. The results were evaluated statistically with Students t-test, chi(2)-test, one-way analysis of variance, and stepwise regression analysis. RESULTS In this study, frequency of PAI-1 gene 4G5G genotype was found to be low both in patients and controls. PAI-1 enzyme activities were significantly increased in acute stroke patients compared to controls. Although PAI-1 gene 4G5G genotype frequencies were low, the patients carrying this allele had highest plasma PAI-1 enzyme activity; likewise, although PAI-1 gene 4G4G genotype frequencies were high, the patients carrying this allele had lowest plasma PAI-1 enzyme activities. Homocysteine levels had a positive effect of 65% on plasma PAI-1 enzyme activities. CONCLUSION Consequently, in this study, we may assert that PAI-1 gene, 4G4G and 5G5G genotypes, PAI-1 activity, and homocysteine level determination are significant criteria for identifying patients who are likely to develop stroke; on the other hand, a direct relation does not exist between gene polymorphism and enzyme activity.

Are the angiotensin-converting enzime gene and acticity risk factors for stroke?

Stroke is a multifactorial disease in which genetic factors play an important role. This study was carried out to determine angiotensin-converting enzyme (ACE) gene polymorphism in Turkish acute stroke patients and to establish whether there is an association of angiotensin-converting enzyme gene I/D polymorphism with clinical parameters. In this study 185 patients and 50 controls were recruited. We have investigated the association among the allelic distribution of the insertion/deletion (I/D) polymorphism of the ACE gene identified by polymerase chain reaction. Distribution of ACE gene I/D genotypes and allele frequencies in patients were not significantly different from controls. D allele frequencies were 57.8% in patients versus 53.0% in controls and I allele 42.2% versus 47% respectively. History of hypertension, stroke, renal, heart and vessel diseases incidence and age, gender, systolic-diastolic blood pressures and creatinine levels were significantly high in patients. But these results and ACE activities had no significant differences among the ACE genotypes in patients and controls. Our results suggest that the ACE gene polymorphism is not associated with the pathogenesis of stroke in Turkish stroke patients.

The protective effects of vitamin E on urinary bladder apoptosis and oxidative stress in streptozotocin-induced diabetic rats.

OBJECTIVES To determine whether vitamin E has protective effects or not on streptozotocin-induced diabetic rats in diabetic urinary bladder dysfunction, with interrelationships between oxidative stress and apoptosis. METHODS Thirty-two Wistar albino male rats were divided into 4 groups. Group A (n = 8), control; group B (n = 8), diabetic control; group C (n = 8), control + vitamin E; and group D (n = 8), diabetic + vitamin E. Vitamin E was injected 40 mg/kg every other day intraperitoneally for 2 weeks. In the diabetic groups, diabetes was induced by a single intraperitoneal injection of 65 mg/kg of streptozotocin. Apoptosis studies were performed using apoptosis detection kit and the TUNEL (TdT-mediated dUTP nick-end labeling) technique. The levels of glucose, malondialdehyde (MDA), superoxide dismutase, catalase, and glutathione peroxidase were detected in hemolysate. RESULTS It was observed that apoptosis number in urothelial cells of the bladder in diabetic rats increased significantly compared with control and decreased after vitamin E treatment. MDA levels of the diabetic group were significantly higher than those on the control and vitamin E groups. Diabetic + vitamin E group had significantly increased MDA levels compared with control group, although these values were lower than those in the diabetic group. All enzyme activities of the vitamin E group did not differ compared with the control group. In diabetic + vitamin E group, superoxide dismutase and glutathione peroxidase activities were similar to controls. Catalase activity of the diabetic + vitamin E group decreased significantly compared with control, although it was higher than that in the diabetic group. CONCLUSIONS Our study revealed that vitamin E decreases apoptosis and may be protective for uroepithelial cells of diabetic bladder.

Determination of the Relationship Between rs4986790 and rs4986791 Variants of TLR4 Gene and Lung Cancer

Chronic inflammation triggers DNA damage and oncogenic mutations and causes tumor formation and tumor progression. One of the important components of the inflammatory response is Toll-like receptor (TLR) family. The objective of our study is to determine the relationship between rs4986790(+896A/G) and rs4986791(+1196C/T) gene polymorphisms and lung cancer risk. PCR-RFLP technique was carried out to identify the genotypes in 100 control individuals and 160 lung cancer patients. DNA extracted from peripheral blood samples were amplified and digested with NcoI and HinfI then visualized. We did not find any difference between genotype frequencies between controls and patients (p > 0.05) in rs4986790. But a significant difference between control group and patients with lung cancer as for genotype frequencies (χ2 = 4.19, p = 0.041) in rs4986791 variants was found. Our data indicate that any correlation was not found between rs4986790 polymorphism and lung cancer, while a correlation between rs4986791 and lung cancer has been determined and found to be associated with lung cancer risk.

Dopamine D2 receptor gene −141C Insertion/Deletion polymorphism in Turkish schizophrenic patients

Schizophrenia is a chronic and neuropsychiatric disease that affects about 0.5–1% of the world’s population. An increase in dopamine and dopamine D2 receptor (DRD2) gene products has been well described in schizophrenic patients. Several groups have studied the relationship between dopaminergic hyperactivity and cellular communications have obtained discordant results. Studies searching for the relationship between the schizophrenia and DRD2 gene have gained more interest. Our objective was to determine the relationships among schizophrenic symptoms in schizophrenia subtypes and severity of symptoms in terms of DRD2 gene −141C Insertion/Deletion [Ins/Del; I/D] polymorphism by PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) assay method. Genomic DNA was prepared from peripheral blood by using salt extraction method. After amplification of genomic DNA, PCR products were digested with BstNI restriction enzyme for the detection of DRD2 gene −141C Ins/Del polymorphism in 73 schizophrenic patients and 60 healthy control subjects. The allelic frequencies of the DRD2 gene −141C Ins/Del polymorphism in case and control groups were 79.5 and 77.5% for I allele; 20.5 and 22.5% for D allele respectively. There was no significant difference in frequencies of genotypes and alleles between the two groups. In schizophrenic and control subjects, there were no significant relationship in severity of the disease and schizophrenia types among the −141C Ins/Del genotypes and alleles.