Investigation of TLR2 (-196 to -174del) and TLR9 (T-1486C) Gene Polymorphisms Association with Inflammatory Bowel Diseases.

BACKGROUND: Inflammatory bowel diseases (IBD), Crohn's disease (CD), and ulcerative colitis (UC) are diseases that result from the combined effects of a predisposing genetic background and several environmental factors, including smoking. Some genes can influence these diseases through genetic inheritance, and their regulation is explained by gene polymorphism. However, Toll-like receptor (TLR) genes have been identified as susceptibility genes for CD and UC. METHODS: A case-control study was performed on a Turkish population composed of 105 healthy controls and  79 CD, 77 UC patients genotyped by Allele-specific PCR and PCR-RFLP for TLR9 (T-1486C) and TLR 2 (-196 to -174del) gene. Genotype and allele frequencies of TLR9 (T-1486C) and TLR 2 (-196 to -174del) gene polymorphisms compared to allele frequencies in CD and UC patients. RESULTS: No statistically significant findings were found between the CD, UC patients, and the control group in terms of both genotype distributions and allele frequencies for TLR 9 (T-1486C; rs187084) and TLR 2 (-196 to -174del; rs111200466) gene polymorphisms in a Turkish population (P > 0.05). CONCLUSION: No association was found between the TLR2 (rs111200466) and TLR 9 (rs187084) gene polymorphisms among IBD patients and the control groups in the Turkish population.

The MNS16A VNTR polymorphism of the TERT gene in bladder cancer.

OBJECTIVE: Bladder cancer (BC) is a complex disease that has a high morbidity rate. The MNS16A polymorphism in the TERT gene has been indicated to play a role in the presence of various cancer types and multiple tumor populations. In the present study, our goal was to investigate whether the MNS16A (VNTRs) in the TERT gene was associated with bladder cancer. MATERIAL AND METHODS: A total of 70 patients with BC and 120 normal controls were included in the study. The MNS16A (VNTRs) in the TERT gene was amplified using polymerase chain reaction (PCR). The PCR products were visualized on 3% high resolution agarose gel and under a UV light. RESULTS: The MNS16A VNTR-302 allele was found to be the most common allele in both, the patient group (64%) and the control group (62%). The second most common allele was the VNTR-243 allele that occurred at a frequency of around 34% in BC patients and 33% in the controls. VNTR-333 (patient group, 1%; control group, 3%) and VNTR-274 (patient group, 2%; control group, 1%) alleles were reported as the least common alleles in this study. CONCLUSION: When comparing the frequencies of genetic variants between cases and controls, we observed that our findings did not support the hypothesis that the MNS16A VNTR polymorphism of the TERT gene might regulate cancer susceptibility.

T-786C, G894T, and intron 4 VNTR (4a/b) polymorphisms of the endothelial nitric oxide synthase gene in bladder cancer cases.

The aim of the present study was to determine whether endothelial nitric oxide synthase (eNOS) gene polymorphisms play a role in development of bladder cancer in the Turkish population. The study was performed on 75 patients (64 men, 11 women) with bladder cancer and 143 healthy individuals (107 men, 36 women) with any kind of cancer history. Three eNOS gene polymorphisms (T-786C promoter region, G894T and intron 4 VNTR 4a/b) were determined with polymerase chain reaction and restriction fragment lenght polymorphism methods. In our study, GT and TT genotypes for eNOS G894T polymorphism were found to significantly vary among patients with bladder cancer and control group (OR: 0.185, CI: 0.078-0.439, p=0.0001 and OR: 0.324, CI: 0.106-0.990, p=0.026). Also, the frequency of the 894T allele was significantly higher in patients with bladder cancer (51%). No association was identified for eNOS T-786C and intron 4 VNTR 4a/b polymorphisms between patients with bladder cancer and control groups in our Turkish population.

Cytogenetic biomonitoring of carpet fabric workers using micronucleus frequency, nuclear changes, and the calculation of risk assessment by repair index in exfoliated mucosa cells.

The micronucleus (MN) assay in exfoliated buccal cells is a minimally invasive method for monitoring genetic damage in human populations and is used as an indicator of genotoxic exposition, as it is associated with chromosome aberrations. In this study, we evaluated MN frequencies and other nuclear changes (NCs), such as karyorrhexis (KR), karyolysis (KL), broken egg (BE), and binucleus in buccal mucosa cells of 50 carpet fabric workers (25 smokers and 25 nonsmokers) and 50 healthy control subjects (25 smokers and 25 nonsmokers). Microscopic observation of 2000 cells per individual was performed in both workers and control subjects. In both the control group and the exposed group, for each person a repair index (RI) was calculated via the following formula: (KR+KL)/(BE+MN). The results showed a statistically significant increase in the frequency of MN in buccal epithelial cells of exposed group compared with control group. There is a significant difference between worker and control groups (p<0.001) for RI. We believe that the calculation of RI values, in addition to nuclear changes, presents a new approach in risk assessment in relation to occupational exposure.

Assessment of genetic damage in buccal epithelium cells of painters: micronucleus, nuclear changes, and repair index.

The micronucleus (MN) assay in exfoliated buccal cells is a useful and minimally invasive method for monitoring genetic damage in humans exposed to occupational and environmental agents. The MN test is used as an indicator of genotoxic exposition, since it is associated with chromosome aberrations. An increased mutation rate in oral squamous cells, which is indicated by an increased MN frequency, is also related to the development of oral mucosa diseases, such as carcinomas. We evaluated MN frequencies and other nuclear changes (NCs), karyorrhexis (KR), karyolysis (KL), broken egg (BE), and binucleus in buccal mucosa cells of 60 painters (30 smokers and 30 nonsmokers) and 60 healthy control subjects (30 smoker and 30 nonsmoker). Microscopic observation of 3000 cells per individual was performed in both painters and control subjects. In the control group and the exposed group, for each person repair index (RI) was calculated via the following formula: (KR + KL)/(BE + MN). The results showed a statistically significant increase in the frequency of MN in buccal epithelial cells of the exposed group compared with the control group (p < 0.05). Smokers and nonsmokers differed with respect to the incidence of MN and NCs in all groups. In painters, RI was less than that in the control group. There was a significant difference between painters and the control group (p < 0.01) for RI. We believe that determination of RI in addition to NCs and the MN will present a new approach to genotoxicity studies of a population.

Nuclear anomalies in the buccal cells of calcite factory workers.

The micronucleus (MN) assay on exfoliated buccal cells is a useful and minimally invasive method for monitoring genetic damage in humans. To determine the genotoxic effects of calcite dust that forms during processing, MN assay was carried out in exfoliated buccal cells of 50 (25 smokers and 25 non-smokers) calcite factory workers and 50 (25 smokers and 25 non-smokers) age- and sex-matched control subjects. Frequencies of nuclear abnormalities (NA) other than micronuclei, such as binucleates, karyorrhexis, karyolysis and 'broken eggs', were also evaluated. Micronuclei and the other aforementioned anomalies were analysed by two way analysis of covariance. The linear correlations between the types of micronucleus and nuclear abnormalities were determined by Spearman's Rho. There was a positive correlation between micronuclei and other types of nuclear abnormalities in accordance with the Spearman's Rho test. Results showed statistically significant difference between calcite fabric workers and control groups. MN and NA frequencies in calcite fabric workers were significantly higher than those in control groups (p < 0.05). The results of this study indicate that calcite fabric workers are under risk of significant cytogenetic damage.

Nuclear anomalies in the buccal cells of calcite factory workers

The micronucleus (MN) assay on exfoliated buccal cells is a useful and minimally invasive method for monitoring genetic damage in humans. To determine the genotoxic effects of calcite dust that forms during processing, MN assay was carried out in exfoliated buccal cells of 50 (25 smokers and 25 non-smokers) calcite factory workers and 50 (25 smokers and 25 non-smokers) age- and sex-matched control subjects. Frequencies of nuclear abnormalities (NA) other than micronuclei, such as binucleates, karyorrhexis, karyolysis and 'broken eggs', were also evaluated. Micronuclei and the other aforementioned anomalies were analysed by two way analysis of covariance. The linear correlations between the types of micronucleus and nuclear abnormalities were determined by Spearman's Rho. There was a positive correlation between micronuclei and other types of nuclear abnormalities in accordance with the Spearman's Rho test. Results showed statistically significant difference between calcite fabric workers and control groups. MN and NA frequencies in calcite fabric workers were significantly higher than those in control groups (p < 0.05). The results of this study indicate that calcite fabric workers are under risk of significant cytogenetic damage.

The genotoxic effect of the new acaricide etoxazole.

Etoxazole is a member of the diphenyl oxazoline class of insecticide was newly developed for use on pome fruits, cotton and strawberries as a acaricide. In the present study, genotoxic effects of acaricide etoxazole (ETX) (miticide/ovicide) were investigated using chromosome aberration (CA) test, sister chromatid exchange (SCE) test and micronucleus test in human lymphocytes. ETX induced the CAs at all concentrations (5, 10 and 20 microg/ml) for 24 h and also induced the CA at the highest concentration (20 microg/ml) for 48 h only. The inducing the CAs for 48 hours treatment period was dose-dependent. Besides, it induced the SCE at all concentrations and treatment periods in a dose-dependent manner as well. Although, ETX decreased the mitotic index (MI) at all concentration and treatment periods dose-dependently, while it did not decrease the replication index (RI) when compared to the negative and solvent controls. In addition, ETX induced the micronucleus at all concentrations except 5 microg/ml for 48 h. This inducing was in a dose-dependent manner as well. In conclusion, it can be concluded that ETX has a potential genotoxic effects in cultured human peripheral lymphocytes.

Genotoxicity of aspartame.

In the present study, the genotoxic effects of the low-calorie sweetener aspartame (ASP), which is a dipeptide derivative, was investigated using chromosome aberration (CA) test, sister chromatid exchange (SCE) test, micronucleus test in human lymphocytes and also Ames/Salmonella/ microsome test. ASP induced CAs at all concentrations (500, 1000 and 2000 microg/ml) and treatment periods (24 and 48 h) dose-dependently, while it did not induce SCEs. On the other hand, ASP decreased the replication index (RI) only at the highest concentration for 48 h treatment period. However, ASP decreased the mitotic index (MI) at all concentrations and treatment periods dose-dependently. In addition, ASP induced micronuclei at the highest concentrations only. This induction was also dose-dependent for 48 hours treatment period. ASP was not mutagenic for Salmonella typhimurium TA98 and TA100 strains in the absence and presence of S9 mix.