Determination of the Roles of Endothelial Nitric Oxide Synthase 4VNTR (4a/b), G894T, T786C Gene Variations in the Bladder Cancer Development.

The aim of this study is to determine the roles of eNOS gene variations in BCA development. Our study included 91 patients diagnosed with BCA and 91 healthy controls. eNOS 4VNTR (4a/b), T786C and G894T gene variations genotype distributions were determined by PCR and RFLP methods. The significant difference was determined between these groups in terms of eNOS T786C and eNOS G894T gene variations genotype distributions (p < 0.05). TT genotype for G894T gene variation and CC genotype for T786C gene variation were detected higher in patients. The CC genotype of T786C gene variation was detected significantly higher in male patients than in male controls (p < 0.05). In addition, aa-TT, ab-TT, bb-TT haplotypes of 4VNTR (4a/b)-G894T gene variations, aa-CC, ab-CC, bb-CC haplotypes of 4VNTR (4a/b)-T786C gene variations and TT-TT, TT-CC, TT-CT, GG-CC, GT-CC haplotypes of G894T-T786C gene variations were observed in patient group more than control group. The significant difference was detected between these groups in terms of eNOS (G894T-T786C) haplotypes (p < 0.05). In our study, eNOS T786C and eNOS G894T gene variations were determined important genetic risk factor in the Thrace population of Turkey.

Investigation of the Roles of MTHFR (C677T and A1298C) and MMP-2 (-1306C>T ) Variations in Bladder Cancer Development.

OBJECTIVE: Bladder cancer is a complex malignancy and has been associated with high morbidity. Since susceptibility to bladder cancer development differs between individuals, determining the roles of MTHFR and MMP-2 gene variations associated with this cancer is important for analyzing differences in individual susceptibility. In this study, we aimed to investigate the role of MTHFR and MMP-2 gene variations in the development of bladder cancer in the Thrace region of Turkey. MATERIALS AND METHODS: One hundred seventy-nine blood samples were collected, including 98 patients with bladder cancer and 81 healthy controls. DNA extraction was carried out with blood samples. Polymerase chain reaction-restriction fragment length polymorphism was applied to detect MTHFR C677T (rs 1801133), MTHFR A1298C (rs 1801131), and MMP-2 (-1306C>T) (rs 243865) gene variants. RESULTS: For the MTHFR A1298C gene variation, CC genotype was the genetic risk factor (P=.0001), while AC genotype was the protective factor (P< .0001) in the development of bladder cancer. For the MMP-2 (-1306C>T) gene variation, TT genotype (P < .0001) and T allele (P=.0006) were genetic risk factors, while AC genotype (P=.0009) was the protective factor in the development of bladder cancer. For C677T/A1298C gene variations, CC-CC combined genotype was the genetic risk factor (P=.009), while CT-AC and CC-AC combined genotypes were potential protective biomarkers (P=.013 and P < .001, respectively). CONCLUSION: In our study, TT genotype and T allele were determined as genetic risk factors for MMP-2 (-1306C>T) gene variation. For C677T/A1298C gene variations, CC- CC combined genotype was detected as the genetic risk factor in the development of bladder cancer.

Investigation of Relationship Between Small Noncoding RNA (sncRNA) Expression Levels and Serum Iron, Copper, and Zinc Levels in Clinical Diagnosed Multiple Sclerosis Patients.

In our study, we aimed to investigate the relationship between microRNA (miRNA) expression levels and serum iron (Fe), copper (Cu), and zinc (Zn) levels in Multiple sclerosis (MS) patients. Total RNA was isolated from peripheral venous blood containing ethylenediaminetetraacetic acid (EDTA) of MS patients and controls. Total RNA was labeled with Cy3-CTP fluorescent dye. Hybridization of samples was performed on microarray slides and arrays were scanned. Data argument and bioinformatics analysis were performed. Atomic absorption spectrophotometer method was used to measure serum Fe, Cu, and Zn levels. In our study, in bioinformatics analysis, although differently expressed miRNAs were not detected between 16 MS patients and 16 controls, hsa-miR-744-5p upregulation was detected between 4 MS patients and 4 controls. This may be stem from the patient group consisting of MS patients who have never had an attack for 1 year. Serum iron levels were detected significantly higher in the 16 MS patients compared to the 16 controls. This may be stem from the increase in iron accumulation based on inflammation in MS disease. According to the findings in our study, hsa-miR-744-5p upregulation has been determined as an early diagnostic biomarker for the development together of insulin resistance, diabetes mellitus associated with insulin signaling, and Alzheimer's diseases. Therefore, hsa-miR-744-5p is recommended as an important biomarker for the development together of diabetes mellitus, Alzheimer's disease, and MS disease. In addition, increased serum Fe levels may be suggested as an important biomarker for neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, and MS disease.

Investigation of roles of IL-8 (+ 781 C/T) and MMP-2 (-735 C/T) gene variations in early diagnosis of bladder cancer and progression.

BACKGROUND: The aim of our study is to investigate the roles of IL-8 (+ 781 C/T) and MMP-2 (-735 C/T) gene variations in early diagnosis and progression of BCA. METHODS: Polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) methods were used to determine the genotype distributions of IL-8 (+ 781 C/T) and MMP-2 (-735 C/T) gene variations. RESULTS: In our study, the genotype distributions of IL-8 (+ 781 C/T) and MMP-2 (-735 C/T) gene variations were not found to be significantly different between the patient and control groups. In addition, C and T allele frequencies for these gene variations were not different from the Hardy-Weinberg distribution in patient and control groups. However, when the combined genotype analyzes for these gene variations were evaluated, CC-CC and CT-CC combined genotypes for + 781 C/T / -735 C/T gene variations were observed significantly more in the patient group compared to other genotypes. CONCLUSION: Although IL-8 (+ 781 C/T) and MMP-2 (-735 C/T) gene variations were not found to be genetic risk factors in the Thrace population in our study, CC-CC and CT-CC combined genotypes were determined as genetic risk factors for BCA susceptibility. The combined genotypes obtained as a result of the combined genotype analysis of these genetic variations that are effective in tumor progression may be considered to be important biomarkers for the early diagnosis and progression of BCA.

CRISPR/Cas9 Mediated Therapeutic Approach in Huntington's Disease.

The pathogenic mechanisms of these diseases must be well understood for the treatment of neurological disorders such as Huntington's disease. Huntington's Disease (HD), a dominant and neurodegenerative disease, is characterized by the CAG re-expansion that occurs in the gene encoding the polyglutamine-expanded mutant Huntingtin (mHTT) protein. Genome editing approaches include zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and Clustered Regularly Interspaced Short Palindromic Repeats/Caspase 9 (CRISPR/Cas9) systems. CRISPR/Cas9 technology allows effective gene editing in different cell types and organisms. Through these systems are created isogenic control of human origin induced pluripotent stem cells (iPSCs). In human and mouse models, HD-iPSC lines can be continuously corrected using these systems. HD-iPSCs can be corrected through the CRISPR/Cas9 system and the cut-and-paste mechanism using isogenic control iPSCs. This mechanism is a piggyBac transposon-based selection system that can effectively switch between vectors and chromosomes. In studies conducted, it has been determined that in neural cells derived from HD-iPSC, there are isogenic controls as corrected lines recovered from phenotypic abnormalities and gene expression changes. It has been determined that trinucleotide repeat disorders occurring in HD can be cured by single-guide RNA (sgRNA) and normal exogenous DNA restoration, known as the single guideline RNA specific to Cas9. The purpose of this review in addition to give general information about HD, a neurodegenerative disorder is to explained the role of CRISPR/Cas9 system with iPSCs in HD treatment.

The roles of MTHFR (C677T, A1298C) and MGP (G-7A, T-138C) gene variations in development of diabetic nephropathy in patients with type 2 diabetes mellitus.

Background: The goal of this study is to determine the role of MTHFR (C677T, A1298C) and MGP (G-7A, T-138C) gene variations in DN development. Methods: There were 61 DN patients and 55 healthy controls in this study. The genotype distributions of these gene variations were determined using PCR and RFLP methods. Results: According to our study, there was no significant relationship between these gene variations and DN (p > 0.05). The allele frequencies of the MTHFR C677T gene variation in the control group were found significantly different from the Hardy-Weinberg distribution (p < 0.05). According to combined genotype analysis, GA-TT combined genotype of MGP (G-7A/T-138C) gene variations was observed significantly more in the patient group with DN. The GA-TT combined genotype of MGP (G-7A/T-138C) gene variations was differ significantly between these groups (OR: 2.359, %95 CI: 1.094-5.087, p = 0.028). Conclusion: In our study, although MTHFR and MGP gene variations were not risk factors, the GA-TT combined genotype of the MGP (G-7A/T-138C) gene variation was detected as an important risk factor for DN.

Investigation of the relationship between MTRR A66G, MTR A2756G gene variations and cell anomalies in early diagnosis and progression of bladder cancer.

BACKGROUND: The aim of this study is to investigate the relationship between MTRR A66G, MTRA2756G gene variations and cell anomalies in the early diagnosis and progression of bladder cancer. METHODS: PCR and RFLP methods were used to determine the genotype distributions of MTRR A66G and MTR A2756G gene variations. Peripheral smear preparations prepared from blood samples were fixed with methanol fixative and stained histochemically. Cellular morphological evaluations were made under the light microscope. RESULTS: In our study, AA-GG haplotype was observed significantly more in the patient group than control group (OR: 3.304, 95% CI: 1.023-10.665, p = 0.046). The significant increase was determined in terms of histological damage parameters in the patient group compared to the control group (p < 0.05). For multiple vacuoles damage parameter (mild score), AA genotype of MTR A2756G gene variation was significantly different compared to AA genotype of MTRR A66G gene variation (OR: 0.211, 0.049-0.912, p = 0.037). AA genotype of MTR A2756G gene variation was observed more than AA homozygous genotype of MTR A66G gene variation for giant platelets with different sizes damage parameter (mild score) (OR: 0.062, 0.017-0.228, p < 0.001). CONCLUSIONS: In conclusion, in Thrace population, AA genotype of the MTR A2756G gene variation was significantly higher than the AA homozygous genotype of the MTR A66G gene variation as a genetic risk factor for the multiple vacuoles damage parameter. In addition, AA genotype of MTR A2756G gene variation was determined as a genetic risk factor for giant platelets with different sizes damage parameter.

Investigation of the Relationship Between IL-18 (- 607 C/A), IL-18 (- 137 G/C), and MMP-2 (- 1306 C/T) Gene Variations and Serum Copper and Zinc Levels in Patients Diagnosed with Chronic Renal Failure.

The aim of this study is to investigate the relationship between IL-18 (- 607 C/A), IL-18 (- 137 G/C), and MMP-2 (- 1306 C/T) gene variations and serum trace element levels in patients diagnosed with CRF. Genotype distributions of IL-18 (- 607 C/A, - 137 G/C) gene variations were determined by polymerase chain reaction (PCR) method. PCR-restriction fragment length polymorphism (RFLP) methods were used to determine the MMP-2 (- 1306 C/T) gene variation genotype distributions. Serum trace element levels were determined by atomic absorption spectrophotometer method. A significant difference was found between the CRF patient and healthy control groups in terms of genotype distributions of IL-18 (- 607 C/A) and MMP-2 (- 1306 C/T) gene variations (p < 0.05). The significant difference was found between the patient and control groups in terms of serum copper and zinc levels and copper/zinc ratio (p < 0.05). The significant difference was detected between patient and control groups in terms of copper and zinc levels and copper/zinc ratio according to IL-18 (- 607 C/A), IL-18 (- 137 G/C), and MMP-2 (- 1306 C/T) gene variations and genotype distributions (p < 0.05). In addition, significant difference was determined in terms of serum copper and zinc levels and copper/zinc ratio according to haplotypes of IL-18 (- 607 C/A), IL-18 (- 137 G/C), and MMP-2 (- 1306 C/T) gene variations between patient and control groups (p < 0.05). In conclusion, evaluation of IL-18 (- 607 C/A, - 137 G/C) and MMP-2 (- 1306 C/T) gene variations and serum trace element levels together is extremely important in terms of obtaining important biomarkers in CRF early diagnosis and progression.

Investigation of the relationship between MMP-1 (- 1607 1G/2G), MMP-3 (- 1171 5A/6A) gene variations and development of bladder cancer.

BACKGROUND: Chronic inflammation is an important risk factor in the development of bladder cancer. It may stimulate growth and metastasis of cancer cells. The inflammatory process includes MMP activities and expression. MMP activation can be stimulated by various inflammatory cells. Pathological processes such as bladder cancer may occur due to imbalance in MMP activities. In our study, we aimed to determine the relationship between MMP-1, MMP-3 gene variations associated with chronic inflammation and the bladder cancer development. METHODS: Our study was carried out with 89 bladder cancer patients and 78 healthy controls. PCR-RFLP methods were applied to determine MMP-1 and MMP-3 gene variations genotype distributions. RESULTS: 1G/1G homozygous and 1G/2G heterozygous genotypes of MMP-1 gene variation were determined more in patients than controls. The 5A/5A homozygous and 5A/6A heterozygous genotypes of the MMP-3 gene variation were detected more in patients than controls. The significant difference was detected in terms of genotype distributions of MMP-1 and MMP-3 gene variations between these groups (p < 0.05). In addition to, the most common haplotype in the patient group were detected as 1G/2G-5A/6A (20.22%). CONCLUSION: In this study, MMP-1 and MMP-3 gene variations were determined as possible genetic risk factors for bladder cancer development in the Thrace population.

Investigation of the relationship between GSTM1 gene variations and serum trace elements, plasma malondialdehyde levels in patients with colorectal cancer.

BACKGROUND: The aim of this study is to investigate of the relationship between GSTM1 gene variations and serum trace elements, plasma malondialdehyde levels in patient with colorectal cancer. Mateials and Methods. Genotype distributions of GSTM1 gene variations were determined using real-time polymerase chain reaction method. Serum trace element levels were determined using atomic absorption spectrophotometer method and plasma MDA levels were measurement by spectrophotometric method. RESULTS: Serum Cu levels, plasma MDA levels and Cu/Zn ratio were determined significantly higher in the group of CRC patient carrying the GA heterozygous genotype of the GSTM1 (rs 112,778,559) gene variation compared to healthy controls (p < 0.05). Serum Cu, Zn levels, plasma MDA levels and Cu/Zn ratio were determined significantly higher in patients carrying GG homozygous genotype of the GSTM1 (rs 112778559) gene variation compared to healthy controls carrying same genotype (p < 0.05). Serum Cu, Zn levels, plasma MDA levels and Cu/Zn ratio were determined significantly higher in the group of CRC patient carrying the GG homozygous genotype of the GSTM1 (rs 12068997) gene variation compared to healthy controls (p < 0.05). On the other hand, serum Se levels were detected significantly lower in CRC patients carrying GA heterozygous and GG homozygous genotypes for GSTM1 (rs 112,778,559) and (rs 12,068,997) gene variations compared to healthy controls (p < 0.05). CONCLUSION: In our study, the evaluation of serum Cu, Zn and Se trace element levels and plasma MDA levels according to GSTM1 gene variations genotype distributions were enabled to obtain important biomarkers in terms of CRC development and progression.

Investigation of the roles of IL-18 (-607 C/A) and IL-18 (-137 G/C) gene variations in bladder cancer development: case-control study.

BACKGROUND: The purpose of our study is to investigate the roles of IL-18 gene variations in bladder cancer development in Thrace population of Turkey. METHODS: This study was carried out with 103 bladder cancer patients and 81 healthy controls. Genotype distributions of IL-18 (-137 G/C) and IL-18 (-607 C/A) gene variations were determined using polymerase chain reaction (PCR) method. RESULTS: The CC homozygous genotype for IL-18 (-607 C/A) gene variation was significantly higher in patients with bladder cancer compared to healthy controls (OR 0.345, 95% Cl 0.186-0.639, p = 0.001). Besides this, allele frequencies of IL-18 (-137 G/C) and IL-18 (-607 C/A) gene variations in patient with bladder cancer and healthy control groups were significantly different from the Hardy-Weinberg distribution (p < 0.05). For IL-18 (-137 G/C) and IL-18 (-607 C/A) gene variations, significant difference was determined between the bladder cancer patient and healthy control groups in terms of GC-CA (OR 0.381, 95% Cl 0.203-0.714, p = 0.002), GC-CC (OR 2.147, 95% Cl 1.013-4.550, p = 0.043), GG-AA (OR 0.431, 95% Cl 0.365-0.509, p = 0.049), and GG-CC (OR 2.476, 95% Cl 1.177-5.208, p = 0.015) haplotypes. CONCLUSION: In our study, CC genotype of IL-18 (-607 C/A) gene variation was determined as genetic risk factor for bladder cancer development. In bladder cancer patient and healthy control groups, G and C allele frequencies of IL-18 (-137 G/C) gene variation, and C and A allele frequencies of IL-18 (-607 C/A) gene variation were determined significantly different from the Hardy-Weinberg distribution.

Investigation of The Relationship Between IL-18 (-607 C/A), IL-18 (-137 G/C) Gene Variations and Ischemic Stroke Disease Development in Thrace Region of Turkey.

BACKGROUND: Ischemic stroke is a clinical condition characterized by focal or global cerebral dysfunction resulting from inhibition of brain blood flow. Genetic factors play an important role in the pathogenesis of ischemic stroke. As a result of IL-18 (-607 C/A, -137 G/C) gene variations, it is thought that binding of transcription factors may be affected and IL-18 mRNA expression can be modulated. Therefore, the purpose of our study is to investigate the roles of IL-18 (-607 C/A), IL-18 (-137 G/C) gene variations in the development of ischemic stroke in Trakya Region of Turkey. METHODS: Our study was performed with 90 ischemic stroke patients and 89 healthy controls. Genotype distributions of IL-18 (-607 C/A, -137 G/C) gene variations were determined using polymerase chain reaction (PCR) method. RESULTS: GC genotype and CA genotype of IL-18 (-137 G/C) and IL-18 (-607 C/A) gene variations were determined higher significantly in patent group as compared with other genotypes. However, the statistically significant difference was not determined between patients with ischemic stroke and healthy control groups in terms of IL-18 (-137 G/C) and IL-18 (-607 C/A) gene variations (p > 0,05). Allele frequencies of IL-18 (-137 G/C) and IL-18 (-607 C/A) in patient and control groups were significantly different from the Hardy-Weinberg distribution (p < .001 for all). CONCLUSION: Although these gene variations' genotype distributions were not determined as a genetic risk factor for the development of ischemic stroke, allele frequencies of IL-18 (-137 G/C) and IL-18 (-607 C/A) in patient and control groups were significantly different from the Hardy-Weinberg distribution.

The association of Intron 4 VNTR and Glu298Asp polymorphisms of the nitric oxide synthetase 3 gene and vasculogenic erectile dysfunction in Turkish men.

Several studies have focused on the impaired role of endothelial nitric oxide synthase (NOS3) gene polymorphism and its association to erectile dysfunction (ED). However, currently controversial results have been reported due to their significant heterogeneity. The present study aimed to assess the genotypic distribution and the allelic frequency of Intron 4 VNTR and Glu298Asp gene polymorphisms in vasculogenic ED patients compared to healthy controls of a specific region of Turkey. A total of 75 patients with ED (median age: 56, IQR:10.5) and 75 healthy controls (median age: 56, IQR:10.5) were prospectively analyzed. All subjects were equally evaluated by the same physician with detailed history-taking, physical examination, International Index of Erectile Function (IIEF) questionnaire, and blood tests (incl. glucose, testosterone, triglyceride and total cholesterol level). Those with an IIEF score under 26 were considered to have ED, by classifying them according to their scores as mild (22-25), moderate (11-21) and severe (1-10) ED. Color doppler ultrasonography was carried out in patients with an IIEF score <22. Hypertension, diabetes mellitus, coronary artery disease, and smoking status were significantly associated with the ED group compared to control subjects with p values of <0.001, <0.001, 0.002 and <0.001, respectively. Overall genotype frequencies was 47 (31%) a/a, 22 (15%) a/b, 82 (55%) b/b for Intron 4 VNTR and 56 (37%) GG, 78 (52%) GT, 16 (11%) TT for the Glu298Asp polymorphism. The frequencies of Intron 4 VNTR a/a allele and Glu298Asp GT allele were associated with severe ED, while a/b and TT were associated with moderate or mild, and b/b and GG were associated with no ED. In contrast to Glu298Asp, statistically significant differences in genotypic frequencies of Intron 4 VNTR gene polymorphism between ED and control subjects was established. Abbreviations: NO: nitric oxide, NOS: nitric oxide synthase, NOS3: endothelial nitric oxide synthase, NOS2: inducible nitric oxide synthase, NOS1: neuronal nitric oxide synthase, HT: hypertension, DM: diabetes mellitus, CAD: coronary artery disease, ED: erectile dysfunction, IIEF: international index of erectile function, VNTR: variable number of tandem repeats, CDU: color doppler ultrasonography, PCR: polymerase chain reaction.