Sacide Pehlivan

Investigation of TNF-alpha, TGF-beta 1, IL-10, IL-6, IFN-gamma, MBL, GPIA, and IL1A gene polymorphisms in patients with idiopathic thrombocytopenic purpura

Immune thrombocytopenic purpura (ITP) is an autoimmune disease characterized by the presence of autoantibodies developing against thrombocyte membrane glycoproteins (GPs), such as GPIIa/IIIa and GPIb/IX. Single nucleotide polymorphisms (SNPs) of inflammatory cytokine genes were investigated in 71 patients with chronic ITP and 71 healthy controls, and they were compared with the clinical parameters. The polymorphisms in the SNPs were investigated with the polymerase chain reaction, polymerase chain reaction with sequence specific primer, and polymerase chain reaction-restriction fragment length polymorphism methods. It was found that the high expression of TNF-alpha (−308) AG phenotype significantly increased in cases with ITP (odds ratio, OR: 0.318, 95% confidence intervals, CI: 0.103–0.987, p < 0.05). TT genotype in TGF-beta 1 (codon 10) significantly decreased in ITP in comparison with the controls (OR: 0.342, 95% CI: 0.149–0.787, p = 0.016). IFN-gamma (+874) TT genotype was detected to be high in cases with ITP (OR: 3.301, 95% CI: 1.400–7.784, p < 0.05), whereas AA genotype was found to be significantly lower (OR: 4.993, 95% CI: 1.586–15.721, p < 0.05). MBL (codon 54) BB genotype (OR: 1.164, 95% CI: 1.059–1.279, p < 0.05) and IL1A A1/A2 genotype (OR: 0.249, 95% CI: 0.076–0.815, p < 0.05) were found to be significantly higher in cases with ITP than in healthy controls. TNF-alpha (−308) AG phenotype was detected to be significantly higher in steroid-refractory and splenectomized cases at the end of the first year than in the steroid–responsive (complete response (CR) and remission (R)) cases (OR: 4.137, 95% CI: 1.156–14.807, p < 0.05). When we compared the cases, from whom we obtained a CR at their first steroid response, with 12 cases, who entered R but from whom we could not obtain any CR, the frequencies of IFN-gamma (+874) AA genotype were found as 12 (20.3%) and 6 (50%) (OR: 0.082, 95% CI: 0.009–0.793, p < 0.05). MBL (codon 54) AB genotype was detected to be significantly higher in CR patients than in R cases (OR: 1.273, 95% CI: 1.110–1.459, p < 0.05). With these findings, it was found that TNF-alpha/AG, TGF-beta 1/TT, IFN-gamma/TT, MBL/BB, and IL-1RA A1/A2 genotypes were detected as the genes of susceptibility to ITP, while TNF-alpha/AG, IFN-gamma/AA, and MBL/AB genotypes might be important in response to steroid treatment.

Association of macrophage migration inhibitory factor gene promoter polymorphisms with multiple sclerosis in Turkish patients.

Elevated levels of macrophage migration inhibitory factor (MIF) have been observed in the cerebrospinal fluid of patients with multiple sclerosis. This study was designed to determine if MIF gene polymorphisms are associated with multiple sclerosis and disease severity. In total, 120 relapsing-remitting patients with multiple sclerosis and 120 control subjects were enrolled in the study. There was a statistically significant increase in the MIF −173 CC genotype in patients with multiple sclerosis compared with the control subjects. The MIF −794 6/7 genotype had a significantly lower progression index compared with MIF −794 6/6. Patients with the MIF −173 CC genotype had a significantly lower age of disease onset compared with those with the MIF −173 CG and MIF −173 GG genotypes. Additionally, patients with the MIF −794 5/6 genotype had a significantly later age of disease onset. This study indicates that the MIF −173 CC genotype may cause susceptibility to multiple sclerosis in the white Turkish population and a younger age of disease onset is associated with this polymorphism.

Gene methylation of SFRP2, P16, DAPK1, HIC1, and MGMT and KRAS mutations in sporadic colorectal cancer

The aim of this study was to investigate the methylation of the SFRP2, P16, DAPK1, HIC1, and MGMT genes, as well as the mutation of amino acid codons 12 and 13 of the KRAS gene in normal and tumor tissue DNA of patients diagnosed with sporadic colorectal cancer (SCRC). The methylation of gene regions and the KRAS mutations of normal (N) and tumor tissue (T) DNA obtained from 17 patients diagnosed with SCRC and 20 healthy controls were investigated using the polymerase chain reaction and reverse-hybridization methods. There was an Asp mutation in four patients, an Asp and Ser mutations in one patient in codon 12 of the KRAS gene, and an Asp mutation in codon 13 in eight patients. Overall promoter methylation (OPM) in the SFRP2 gene was observed in one N and four T, whereas partial promoter methylation (PPM) was observed in two N and five T. OPM in the P16 gene was present in one T. In the DAPK1 gene, OPM existed in seven T and five N, while PPM was present in two N. In the HIC1 gene, OPM was demonstrated in three T, while PPM was noted in two N; however, no methylation existed in N. In the MGMT gene, OPM occurred in five T and two N, and PPM was present in one T. KRAS mutations in Turkish patients with SCRC are similar to those of other population groups. Methylations in the genes, which underwent methylation analysis, were higher in T in comparison with N, and it has been suggested that significant results would be obtained by making a study with a larger population.

Role of cytokine gene (IFN-γ, TNF-α, TGF-β1, IL-6, and IL-10) polymorphisms in pathogenesis of acute rheumatic fever in Turkish children

Acute rheumatic fever (ARF) is a delayed immunologically mediated sequela of throat infection by group A β-hemolytic streptococci. Inflammatory cytokines may play a pathogenic role in ARF. The objective of this study was to investigate the potential associations between interferon (IFN)-γ, interleukin (IL)-6, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β1, and IL-10 gene polymorphisms and childhood ARF. Thirty-eight ARF patients and 40 age- and sex-matched healthy controls were analyzed for eight polymorphisms in five different cytokine genes [IFN-γ (+874), IL-6 (−174), TNF-α (−308), TGF-β1 (+10, +25), and IL-10 (−1082, −819, −592)]. Cytokine genotyping was performed by polymerase chain reaction sequence-specific primer methods. Patients with ARF had significantly higher frequencies of IFN-γ (+874) polymorphism in both TT genotype (p = 0.0002) and T allele (p = 0.0004). No statistically significant differences were observed in genotypes, haplotypes, and allele frequencies of IL-6, TNF-α, TGF-β1, and IL-10 genes between ARF and control groups (p > 0.05). GG genotype frequency of TNF-α gene (low expression) was higher in patients who had previous ARF history (p = 0.006). High expression of TGF-β1 (TT/GG, TC/GG) was more frequent in patients with CRP positivity (p = 0.034). IL-6 CC genotype (low expression) frequency was higher in patients with tricuspid valve insufficiency (p = 0.002), while IFN-γ TT genotype (high expression) frequency was higher in patients with mitral valve prolapse (p = 0.049). Conclusion: High expression of the IFN-γ gene may carry a higher risk for ARF in Turkish children, while IL-6, TNF-α, and TGF-β1 may have an impact in mediating some clinical and laboratory manifestations of the disease.

Analysis of Dopamine D2 Receptor (DRD2) Gene Polymorphisms in Cannabinoid Addicts

Abstract:  The gene encoding the dopamine D2 receptor (DRD2) has been suggested as a candidate gene for substance dependence. In this study, the possible association between Taq1A and Taq1B DRD2 polymorphisms and cannabinoid dependence was investigated. One hundred and twelve cannabinoid addicted and 130 healthy control subjects were included in this study. The Taq1A and Taq1B genotypes were determined in all subjects by polymerase chain reaction. For each polymorphism (A or B), the subjects were categorized into three groups according to their genotype, that is, the subjects with alleles A1/A1, A1/A2, A2/A2; B1/B1, B1/B2, and B2/B2. A significant association was found between Taq1A gene polymorphism and cannabinoid addicts compared to the control subjects. This finding suggests that polymorphism of the Taq1A, but not the Taq1B, may be associated with the susceptibility to cannabinoid dependence. Further clinical studies are required to be carried out for confirmation and evaluation of these findings.

Enzyme levels and G-463A polymorphism of myeloperoxidase in chronic lymphocytic leukemia and multiple myeloma

The aim of this study was to investigate how myeloperoxidase (MPO) G-463A gene polymorphism and enzyme levels varied among patients with chronic lymphocytic leukemia (CLL) and multiple myeloma (MM) and to find the relationship between the MPO gene, enzyme levels, and clinical parameters. We studied the sera from 40 healthy volunteers, patients with CLL (n = 34) and MM (n = 28). In subjects with homozygote GG genotype, MPO levels were higher in the patients with both CLL and MM than in the control group. This difference was statistically significant in patients with CLL. In conclusion, homozygote GG genotype is found to be associated with an increasing amount of serum MPO. In accordance with the results of the study, we assess that the increase in the MPO enzyme level in the patient groups with CLL and MM generated bactericidal effects as well as the increased formation of ROP, thus setting off a pro-cell death pathway and playing a role on the pathogenesis of lymphoproliferative malignancies through this mechanism.

Pregnancy in patients with chronic myeloid leukemia treated with imatinib.

Pregnancy in patients with chronic myeloid leukemia treated with imatinib Mehmet Yilmaz a; Osman Demirhan b; Ercan Kucukosmanoglu c; Mustafa Pehlivan a; Vahap Okan a; Ozcan Balat d; Sacide Pehlivan e a Department of Hematology, School of Medicine, Gaziantep University, Gaziantep, Turkey b Department of Medical Biology and Genetics, Faculty of Medicine, Cukurova University, Adana, Turkey c Departments of Pediatrics, d Obstetrics and Gynecology, e Medical Biology and Genetics, School of Medicine, Gaziantep University, Gaziantep, Turkey

TNF-α promoter polymorphisms in multiple sclerosis: no association with -308 and -238 alleles, but the -857 alleles in associated with the disease in Turkish patients.

Dysregulation in the expression of pro‐ and anti‐inflammatory cytokines is one of the milestones in multiple sclerosis (MS) development and progression. Tumour necrosis factor (TNF‐α), a proinflammatory cytokine is believed to play an important role in MS pathogenesis. The objective of this study is to investigate the association between TNF‐α promoter region (TNF‐α−238, −308 and −857) and susceptibility to MS and clinical course of the disease. Eighty‐six relapsing remitting MS patients and 150 sex‐, age‐ and ethnic‐matched controls were enrolled in the study. Genotyping was performed by PCR‐RFLP method. We observed a statistically significant increase in TNF‐α 857 CC genotype in MS patients than controls (P < 0.001) while TNF‐α 857 CT genotype showed a significant negative correlation with MS patients (P = 0.033). No differences in the distribution of the TNF‐α−238 and −308 alleles were observed. None of the three polymorphisms (−238, −308 and −857) did not show relation with disease duration, Expanded Disability Status Scale or age of onset. On the other hand, significant difference of TNF −857 CC genotype was identified with the low disease index (P = 0.025). Although the study group is small, the results indicate that TNF‐α 857 CC genotype may cause susceptibility to MS in the Turkish population.

Association of macrophage migration inhibitory factor and mannose-binding lectin-2 gene polymorphisms in acute rheumatic fever

BACKGROUND Macrophage migration inhibitory factor and mannose-binding lectin-2 play important roles in the pathogenesis of several acute and chronic inflammatory/autoimmune disorders. The aim of the study was to investigate any possible association between migration inhibitory factor and mannose-binding lectin-2 gene polymorphisms and acute rheumatic fever in children. Material and methods A total of 38 unrelated children with acute rheumatic fever and 40 age- and sex-matched healthy controls were analysed for codon 54 A/B polymorphism in mannose-binding lectin-2 gene and -173 G/C polymorphism in migration inhibitory factor gene by using the polymerase chain reaction method. RESULTS Frequency of BB genotype of mannose-binding lectin-2 gene was higher in the patient group. Interestingly, children with acute rheumatic fever with AA genotype tended to have chorea compared with children with BB genotype. There was a statistically significant increase in frequency of the migration inhibitory factor -173 CC genotype in patients compared with the control subjects. CONCLUSION The present study is the first to investigate the mannose-binding lectin-2 gene polymorphism in children with acute rheumatic fever. BB genotype of mannose-binding lectin-2 (codon 54) and CC genotype of migration inhibitory factor (-173) may have a role in the immunoinflammatory process of acute rheumatic fever.

Genetic Variation of Myeloperoxidase Gene Contributes to Preeclampsia: A Preliminary Association Study in Turkish Population

Objective. To examine MPO gene polymorphisms of women with preeclampsia in Turkish population. Methods. Sixty-one preeclamptic and 61 normotensive women without history of preeclampsia in earlier pregnancies were enrolled in this prospective controlled study. MPO mutations were characterized by PCR-RFLP method. Results. We demonstrated a significant difference in patients with preeclampsia in terms of genotype frequency. Heterozygous carriers of −463A among preeclamptic pregnancies were significantly frequent, whereas rare A/A homozygotes failed to differ from controls. Conclusion. The −463G/A polymorphism of leukocyte MPO could be an intriguing susceptibility factor that modulates an individuals risk of preeclampsia in Turkish population.