Homozygous Mutant Genotype Research Articles

Association of SLC22A1, SLC47A1, and KCNJ11 polymorphisms with efficacy and safety of metformin and sulfonylurea combination therapy in Egyptian patients with type 2 diabetes.

Multidrug and toxin extrusion transporter 1 (MATE1), encoded by the SLC47A1 gene and single nucleotide polymorphisms of organic cation transport 1, may impact metformin's responsiveness and side effects. Inward-rectifier potassium channel 6.2 (Kir 6.2) subunits encoded by KCNJ11 may affect the response to sulfonylurea. This study aimed to evaluate the association between SLC22A1 rs72552763 and rs628031, SLC47A1 rs2289669 and KCNJ11 rs5219 genetic variations with sulfonylurea and metformin combination therapy efficacy and safety in Egyptian type 2 diabetes mellitus patients. This study was conducted on 100 cases taking at least one year of sulfonylurea and metformin combination therapy. Patients were genotyped via the polymerase chain reaction-restriction fragment length polymorphism technique. Then, according to their glycated hemoglobin level, cases were subdivided into non-responders or responders. Depending on metformin-induced gastrointestinal tract side effects incidence, patients are classified as tolerant or intolerant. KCNJ11 rs5219 heterozygous and homozygous mutant genotypes, SLC47A1 rs2289669 heterozygous and homozygous mutant genotypes (AA and AG), and mutant alleles of both polymorphisms were significantly related with increased response to combined therapy. Individuals with the SLC22A1 (rs72552763) GAT/del genotype and the SLC22A1 (rs628031) AG and AA genotypes were at a higher risk for metformin-induced gastrointestinal tract adverse effects. The results implied a role for SLC47A1 rs2289669 and KCNJ11 rs5219 in the responsiveness to combined therapy. SLC22A1 (rs628031) and (rs72552763) polymorphisms may be associated with increased metformin adverse effects in type 2 diabetes mellitus patients.

The Impact of IL-4 on Abortion Rates in Iraqi Women's Last Trimester Pregnancies

The term "abortion" is used to describe the termination of a pregnancy through the surgical expulsion of a developing embryo or fetus. Whole samples were collected from Al-Zahraa Teaching Hospital in Wasit city From mid-October 2022 until mid-October 2023. The total number of samples was 100 divided into two groups: the first group is women who suffer from a miscarriage in the third trimester and the control group .Result: This table shows the interpretation of the results between patients and healthy people, where the homozygous CC genotype significantly decreased comparing with control (64 versus 86%, OR=0.18; p-value =0.03 ), while the CT heterozygous genotype was significantly high with risk in comparison with control (32 versus 8%; OR= 7.3; p-value 0.005). The study of polymorphism of the IL-4 gene and its association with abortion had limited available data. This study is the second to report on the association between polymorphism of IL-4 and abortion. showed a clear difference of the parameter’s levels in abortion study group. Based on the mutant homozygous CC genotype. In the present study, the significant change in IL-4 concentration was observed due to the effect of the polymorphism in cases of abortion. These findings were consistent with a previous study that reported higher concentrations of IL-4 in women with abortion compared to normal women (P-value of 0.001). (Association between maternal circulating IL-4 levels and preeclampsia).

The role of methylenetetrahydrofolate reductase gene polymorphisms hypercoagulable status of coronavirus disease

Abstract Background: Hypercoagulation is a hallmark in coronavirus disease (COVID-19). The activity of the enzyme methylenetetrahydrofolate reductase (MTHFR) determines homocysteine levels, and polymorphisms in the enzyme’s gene can influence the enzyme activity with a consequence of hypercoagulability in patients with COVID-19. Objectives: To investigate the association of two single nucleotide polymorphisms (SNPs) in the MTHFR gene with hypercoagulability status in COVID-19. Materials and Methods: This is a retrospective cross-sectional study, which included 90 patients diagnosed with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with variable severity. Patients were classified according to D-dimer level at admission into two groups: with and without hypercoagulability. Nucleic DNA was extracted from leukocytes and gene fragments corresponding to C677T and A1298C MTHFR gene were amplified and genotyped using allele specific polymerase chain reaction. Results: Hypercoagulation was reported in 42.22% of the patients. The mutant homozygous genotype (TT) was more frequent among hyper - than normocoagulable patients (13.6% vs. 1.92%) with a significant difference (odds ratio [OR] = 9.28, 95% confidence interval [CI] = 1.02–84.78, P = 0.048). Furthermore, T allele was more common among hyper- than normocoagulable patients (28.95% vs. 13.46%) with a significant difference (OR = 2.62, 95% CI = 1.24–5.5, P = 0.012). In contrast, the SNP A1298C had no significant impact. Conclusions: The TT genotype and T allele of C677T polymorphism but not A1298C in cMTHFE gene could be considered a risk factor for the hypercoagulable status in COVID-19.

Polymorphic Analysis of Genes PADI4 (rs2240340, rs1748033) and HLA-DRB1 (rs2395175) in Arthritis Patients in Pakistani Population.

Genes are an important factor for the initiation of any disease. Many genes are associated with rheumatoid arthritis (RA) other than environmental factors. The main objective of the study was to evaluate the association of genes PADI4 (peptidylarginine deiminases 14) (rs2240340, rs1748033) and Human leukocyte antigen class II histocompatibility, D-related beta chain (HLA-DRB1) (rs2395175) polymorphisms in RA patients from Punjab, Pakistan. Blood samples of RA patients were collected from different hospitals of Sargodha. DNA was extracted, followed by PCR. Polymorphic analysis was performed in 300 rheumatoid arthritis patients and 300 healthy controls on PADI4 (rs2240340, rs1748033) and HLA-DRB1 (rs2395175). In PADI4 gene, both homozygous mutant genotype (TT) and heterozygous (CT) of SNP rs2240340 showed significant association by increasing the risk of RA up to twofold (OR 2.55; 95% CI 1.57-4.15; p = 0.0002). In case of rs1748033 polymorphism, homozygous mutant genotype (TT) showed significant association with RA by increasing therisk of disease up to threefold (OR 3.46; 95% CI 1.97-6.07; p = 0.0001), while heterozygous genotype (CT) of the same SNP showed significant association with RA by playing a protective role (OR 0.57; 95% CI 0.36-0.91; p = 0.0197). In HLA-DRB1 gene, homozygous mutant genotype (GG) of SNP rs2395175 showed no significant association with RA, while heterozygous genotype (AG) of the same SNP showed significant association with RA by playing a protective role (OR 0.44; 95% CI 0.27-0.71; p = 0.0009). Highly significance association of genes PADI4 (rs2240340, rs1748033) and HLA-DRB1 (rs2395175) polymorphisms with RA was observed in Pakistani population.

MiRNA-767 and its binding site polymorphism in the mTOR gene act as potential biomarkers for female reproductive cancers.

Aims: The present study aimed to understand the relationship between the mTOR gene SNP (rs2536) and reproductive cancer risk. The expression level of miRNA-767 was also assessed. Methods: 700 tumor samples (300 breast, 200 ovarian and 200 cervical cancers), along with adjacent uninvolved control tissue, were used. rs2536 was screened using Tetra-ARMS PCR and expression level of miRNA-767 was assessed using quantitative PCR. Results: The frequency of the homozygous mutant genotype of rs2536 was observed significantly higher in breast (p<0.04), ovarian (p<0.005) and cervical (p<0.003) cancers. Significant downregulation of miRNA-767 was observed in tumors compared with controls. Conclusion: The present study demonstrates that increased mutant frequency of rs2536 and deregulation of miRNA-767 are associated with increased reproductive cancer risk.

Genetic mutation rs972283 of the KLF14 gene and the incidence of gastric cancer

Introduction and Aim: Genetic factors and family gene clustering constitute an important ratio for gastric cancer. Kruppel Like Factor 14 (KLF14) gene has a carcinogenic role and a clear role in metabolic diseases, but how this gene regulates these metabolic traits is still obscure. Previous studies proposed that the accumulation of single nucleotide polymorphisms (SNPs) in KLF14 may be associated with gastric cancer. The current study aimed to investigate whether single nucleotide polymorphisms (SNP) rs972283 in KLF14 is associated with an increased risk of gastric cancer in the Iraqi population. Materials and Methods: The SNP was genotyped using tetra primer ARMS-PCR in 101 (79 men and 22 women) gastric cancer patients who did not receive chemotherapy, and 80 healthy controls (53 men and 27 women). All patient samples were taken from the Baghdad Hospital of Gastroenterology and Hepatology laboratories. Patient records included age, sex, histological type, and H. pylori infection status Results: The KLF14 rs972283 genotype was significantly different between the gastric cancer and control groups. The heterozygous AG genotype and A mutant allele were significantly higher in gastric cancer patients compared to controls (56.4% vs 38.7%, p&lt;0.01 and 61% vs 40.6%, p&lt;0.01, respectively). In contrast, the GG wildtype genotype and G wildtype allele were significantly higher in controls (40% vs 11%, p&lt;0.01 and 59.4% vs 39%, p&lt;0.01, respectively). The AA homozygous mutant genotype also showed a weak correlation with increased gastric cancer risk. These results indicate the A allele is a risk factor while the G allele has a protective effect for gastric cancer. Conclusion: the KLF14 polymorphism rs972283 exhibits a significant association with gastric cancer risk in our Iraqi cohort. The SNP may serve as a useful prognostic marker, pending validation in larger studies.

CYP1A1 gene polymorphism and heavy metal analyses in benign prostatic hyperplasia and prostate cancer: An explorative case-control study.

The prostatic disorder is associated with benign prostatic hyperplasia (BPH) and prostate cancer (CaP). Evidently, prevalent transcription factors and signaling pathways define their relationship. The etiology of the prostatic disorder is multifactorial including heavy metal toxicity like lead (Pb), Cadmium (Cd), and genetic factors. This study elucidates the association between heavy metal toxicity Pb, Cd, and CYP1A1 gene polymorphism with BPH and CaP. a case-control study with (BPH, n = 104), (CaP, n = 58) and (controls, n =107) patients. Heavy metal Pb and Cd estimation by atomic absorption spectrophotometer. The polymorphism of the CYP1A1 T>C (rs4646903) gene was analyzed byPCR-RFLP. Higher levels of Pb and Cd were found in BPH and CaP followed by the control group (P-value: < 0.05). Pb and Cd show a significant correlation among prostate volume in CaP. Additionally, PSA, IPSS score, and pre void volume were positively co-related with Pb in BPH patients. The posthoc test defines the level of Pb and Cd as significantly elevated in the mutant genotype, highest among homozygous mutant genotype of CYP1A1gene among BPH. In CaP, Pb is significantly higher among the homozygous mutant type of CYP1A1 gene. The risk is also influenced by smoking, tobacco, and alcohol. The heavy metal toxicity Pb and Cd were reported to raise the risk of BPH and CaP. However, a person with heavy metal toxicity especially in BPH has a high-risk genetic susceptibility to the CYP1A1 gene in the north Indian population.

IRF4 is required for migration of CD4+ T cells to the intestine but not for Th2 and Th17 cell maintenance.

The transcription factor Interferon Regulatory Factor 4 (IRF4) is central in control of T cell activation and differentiation. Deficiency of IRF4 results in severe immune deficiency and affects maturation and function of most if not all T cell subsets. Here we use mouse infection models for Citrobacter rodentium and Strongyloides ratti to analyze the function of IRF4 in T helper (Th) 17 and Th2 cell responses, respectively. IRF4 deficient mice were impaired in the control of both pathogens, failed to mount Th17 and Th2 cell responses and showed impaired recruitment of T helper cells to the intestine, the infection site of both pathogens. Compromised intestinal migration was associated with reduced expression of the intestinal homing receptors α4β7 integrin, CCR9 and GPR15. Identification of IRF4 binding sites in the gene loci of these receptors suggests a direct control of their expression by IRF4. Competitive T cell transfer assays further demonstrated that loss of one functional Irf4 allele already affected intestinal accumulation and Th2 and Th17 cell generation, indicating that lower IRF4 levels are of disadvantage for Th2 and Th17 cell differentiation as well as their migration to the intestine. Conversion of peripheral CD4+ T cells from an Irf4 wildtype to an Irf4 heterozygous or from an Irf4 heterozygous to a homozygous mutant genotype after C. rodentium or S. ratti infection did not reduce their capacity to produce Th17 or Th2 cytokines and only partially affected their persistence in the intestine, revealing that IRF4 is not essential for maintenance of the Th2 and Th17 phenotype and for survival of these T helper cells in the intestine. In conclusion, we demonstrate that the expression levels of IRF4 determine Th2 and Th17 cell differentiation and their intestinal accumulation but that IRF4 expression is not crucial for Th2 and Th17 cell survival.

#3145 ASSOCIATION OF ENDOTHELIAL NITRIC OXIDE SYNTHASE GENE POLYMORPHISM WITH ENDOTHELIAL DYSFUNCTION IN DIABETIC NEPHROPATHY AMONG AZERBAIJANIS

Abstract Background and Aims Vascular endothelial dysfunction resulting from impaired nitric oxide synthase (NOS) activity in the endothelial cells of blood vessels has been suggested as playing an important role in the pathogenesis of Diabetic nephropathy (DN). It regulates endothelial function and maintains endothelial-dependent vasodilation in multiple organs, including the kidney. The study was carried out to investigate the association of T-786C single nucleotide polymorphisms with the flow mediated dilation (FMD) in DN among type 2 diabetic subjects. Method 90 patients (45 male and 45 female) with diabetic nephropathy were recruited from Nephrology and Endocrinology departments of the Azerbaijan Medical University hospital. As a control group the results were compared with the findings in 100 healthy Azerbaijani individuals (50 males and 50 females). The study was approved by ethics committee and informed consent was obtained from all patients in written form. Brachial endothelial function was determined by brachial artery vasodilator response or brachial FMD in the dominant arm according to previously validated techniques. T786C polymorphism was detected using polymerase chain reaction-restriction fragment length polymorphism analysis. Results The results showed significant differences between DN group and control group regarding the genotype and allele distributions of the T786C single nucleotide polymorphisms. The CC genotype was significantly more frequent in diabetics nephropathy patients than in control group (42% vs. 8%, p = 0.01). FMD in allele homozygote T786C(TT), T786C(CC) and heterozygote T786C(CT) were respectively 13.5 ±0.16%, 12.3±0.19%,13.9 ±0,28%. In the study groups, Azerbaijani patients showed dominance of the C allele and a mutant homozygous genotype CC of the T786C gene of eNOS. Subjects with the 786C(CC) (n = 48) showed significantly reduced FMD with those with T786C (TT) (n = 7) (p = 0.021) and T786C (CT) (n = 35) (p = 0.023), whereas there was no significant difference in endothelial-independent vasodilation between these groups. Although subjects with homozygote T786C(TT), did not show significantly reduced levels of FMD. Who were carriers of the T786C allele demonstrated markedly reduced levels of FMD compared with noncarriers (p = 0.019). Conclusion The data suggest that in diabetic nephropathy, the eNOS T786C allele may be involved in endothelial dysfunction. Azerbaijani patients showed dominance of the C allele and a mutant homozygous genotype CC of the T786C gene of eNOS.

Polymorphisms of the interleukin-6 (IL-6) gene contribute to cervical cancer susceptibility in Bangladeshi women: A case-control study.

Cervical cancer is characterized by abnormal cell growth in the lining of cervix and it is the second major cause of cancer-related deaths among females in Bangladesh. Interleukin-6 (IL-6) is a multifunctional cytokine that has been heavily linked with cervical cancer. Our aim was to investigate the association of two promoter single-nucleotide polymorphisms (SNPs) of IL-6 (rs1800795 and rs1800797) with the susceptibility of cervical cancer in Bangladeshi women. DNA was extracted from venous blood samples from cervical cancer patients (n = 126) and healthy controls (n = 120). Polymerase chain reaction-restriction fragment length polymorphismwas used for genotyping of the selected SNPs. Logistic regression was performed to calculate the odds ratio (OR)with 95% confidence interval (CI) and p values. We found a significant association between rs1800795 and rs1800797 polymorphisms and cervical cancer. For, rs1800795 (G > C) the GC heterozygous genotype (OR = 2.80, 95% CI = 1.55-5.07, p = 0.0007) and CC mutant homozygous genotype (OR = 3.5, 95% CI = 1.29-9.51, p = 0.014) conferred an increased risk of cervical cancer. In case of rs1800797 (G > A) polymorphism, the AG heterozygous genotype (OR = 6.94, 95% CI = 3.76-12.81, p < 0.0001) and AA mutant homozygous genotype (OR = 3.88, 95% CI = 1.12-13.51, p = 0.0332) also exhibited an elevated risk of cervical cancer. Use of contraceptives was found as risk factor and patients who smoke were carriers of both the risk alleles and thus had an increased risk of cervical cancer. Our findings suggest that polymorphism of rs1800795 and rs1800797 of the IL-6 gene play a significant role in cervical cancer susceptibility in Bangladeshi women.

Association of Methylenetetrahydrofolate Reductase Gene Polymorphism in Mothers With Adverse Clinical Outcomes in Neonates.

The presence of polymorphic methylenetetrahydrofolate reductase(MTHFR) in mothers poses a risk for numerous detrimental outcomes in neonates. The present study investigated the association of maternal MTHFR A1298C and C677T single nucleotide polymorphisms (SNPs) with the clinical outcomes in their neonates. The cross-sectional study included 60 mothers and their neonates. Blood samples from mothers were analyzed for MTHFR A1298C and C677T SNP genotyping by real-time polymerase chain reaction. Clinical details of mothers and neonates were documented. Study groups were stratified based on wild, heterozygous, and mutant genotypes for the respective polymorphisms observed in mothers. Multinomial regression was applied for the association, followed by gene model formulation to estimate the impact of the genetic variants on the outcomes. The frequency percentages of mutant CC1298 and TT677 genotypes were 25% and 8.06%, respectively, and the mutant allele frequencies (MAF) were 42.5% and 22.5%. Percentages of adverse outcomes such as intrauterine growth restriction, sepsis, anomalies, and mortality were higher in neonates born to mothers with homozygous mutant genotypes. Maternal C677T MTHFR SNPs revealed a significant association with neonatal anomalies (p = 0.001). The multiplicative risk model depicted OR (95% CI) for CT vs. CC+TT as 3.0 (95% CI: 0.66-13.7), and for TT vs. CT+CC was 15 (95% CI: 2.01-112.12). The C677T SNP in mothers predicted a dominant model for neonatal death (OR (95% CI): 5.84 (0.57-60.03), p = 0.15), whereas the A1298C reported recessive model for 1298CC mothers (OR (95% CI): 11 (1.05-115.5), p = 0.02). Both the genotypes assumed a recessive model for adverse neonatal outcomes: OR (95%CI) for CC vs. AA+ACwas 3.2 (0.79-12.9, p = 0.1), and for TT vs. CC+CT was 5.48 (0.57-175.7, p = 0.2). The risk for sepsis in neonates was nearly six times higher in those born from mothers with homozygous CC1298 and TT677 than in the wild and heterozygous variants. Mothers with C677T and A1298C SNPs are highly susceptible to adverse outcomes in their neonates. Hence, screening the SNPs during the antenatal period can purposefully serve as a better predictive marker, following which proper clinical management could be planned.

Analysis of CYP2C19 gene polymorphism and influencing factors of pharmacological response of clopidogrel in patients with cerebral infarction in Zhejiang, China.

Certain genetic and non-genetic factors may cause damaged platelet inhibition by clopidogrel. We aimed to determine the effect of cytochrome P4502C19 (CYP2C19) polymorphism, along with other clinical factors, on the platelet response to clopidogrel in patients with acute ischemic stroke (AIS). A total of 214 patients with AIS receiving clopidogrel at a maintenance dose of 75 mg daily admitted to the Ningbo First Hospital between 1 January 2020, and 31 December 2021, were enrolled. Platelet aggregation analysis was performed to determine clopidogrel resistance. Quantitative real-time polymerase chain reaction (QRT-PCR) was used to determine CYP2C19 genotype. Other laboratory data on complete blood count and biochemical parameters were taken from patient medical files. Among the 214 AIS patients treated with clopidogrel in the Ningbo population, the incidence of clopidogrel resistance was approximately 43.9%, and the distribution of CYP2C19 genotypes was highest for CYP2C19(*1/*2) (43.0%), followed by CYP2C19 (*1/*1) (38.8%). The distribution of alleles *1, *2, *3, and *17 was 62.1, 32.5, 4.9, and 0.5%, respectively. A chi-squared test showed that the gene frequencies of alleles *2 and *3 were significantly higher in the clopidogrel-resistant group than in the clopidogrel-sensitive group (p < 0.001), and a Mann-Whitney U-test showed that high HCY levels were significantly correlated with clopidogrel resistance (p < 0.001). Multi-factor logistic regression analysis demonstrated that mutant heterozygous genotype [OR 2.893; 95% confidence interval (CI) 1.456-5.748; p = 0.002], mutant homozygous genotype (OR 4.741; 95% CI 1.828-12.298; p = 0.001), and high HCY levels (OR 1.209; 95% CI 1.072-1.362; p = 0.002) were significantly associated with clopidogrel resistance. According to our results, carrying the CYP2C19*2/*3 allele and high HCY levels are independent risk factors for clopidogrel resistance after clopidogrel therapy in patients with AIS. These two factors should be considered prior to clopidogrel administration.

The IRF6 AP-2α binding site polymorphism relate to the severity of non-syndromic orofacial cleft of Indonesian patients.

IRF6 AP-2α binding site polymorphism is known as IRF6 rs642961. It has been associated with a nonsyndromic orofacial cleft (NS OFC). This study aimed to determine the IRF6 rs642961 as a risk factor associated with NS OFC and its phenotypes. The case-control design used for 264 subjects consists of 158 NS CLP subjects (42 CU CLP, 34 CB CLP, 33 CLO, 49 CPOs) and 106 healthy controls. The DNA is extracted from venous blood. The segment of IRF6 rs642961 amplified by polymerase chain reaction (PCR) followed by restriction fragment length of polymorphisms (RFLPs) used the MspI digestion enzyme. The qPCR method to identify the mRNA expression levels of the IRF6 gene rs642961 was analyzed by the Livak method. The study results show that in NS CB CLP phenotype as the most severe phenotype of NS OFC, the Odds Ratio (OR) of A mutant allele was 5.094 (CI=1.456-17.820; P=0.011) and the OR of AA homozygous mutant genotype was 13.481 (CI=2.648-68.635; P=0.001). There are different levels of mRNA expression changes from NS OFC and its phenotypes. It is substantial among the 2-ΔΔCt and the group of AA, GA, and GG genotypes (P<0.05); in the NS CPO phenotype, it shows IRF6 mRNA under-expression in GA, AA genotypes while in other phenotypes it shows IRF6 mRNA overexpression. The IRF6 AP-2α binding site polymorphism is strongly associated with the severity of NS OFC, and this polymorphism has a functional role in affecting IRF6 mRNA expression that is variable in each phenotype.

Additive Effects of Drinking Habits and a Susceptible Genetic Polymorphism on Cholesterol Efflux Capacity.

High levels of high-density lipoprotein cholesterol (HDL-C) are not necessarily effective in preventing atherosclerotic cardiovascular disease, and cholesterol efflux capacity (CEC) has attracted attention regarding HDL functionality. We aimed to elucidate whether drinking habits are associated with CEC levels, while also paying careful attention to confounding factors including serum HDL-C levels, other life style factors, and rs671 (＊2), a genetic polymorphism of the aldehyde dehydrogenase 2 (ALDH2) gene determining alcohol consumption habit. A cross-sectional study was performed in 505 Japanese male subjects who were recruited from a health screening program. Associations of HDL-C and CEC levels with drinking habits and ALDH2 genotypes were examined. The genotype frequencies of ALDH2 ＊1/＊1 (homozygous wild-type genotype), ＊1/＊2 and ＊2/＊2 (homozygous mutant genotype) were 55%, 37% and 8%, respectively. Both HDL-C and CEC levels were higher in ALDH2 ＊1/＊1 genotype carriers than in ＊2 allele carriers. Although HDL-C levels were higher in subjects who had a drinking habit than in non-drinkers, CEC levels tended to be lower in subjects with ≥ 46 g/day of alcohol consumption than in non-drinkers. Furthermore, CEC levels tended to be lower in ALDH2 ＊1/＊1 genotype carriers with a drinking habit of ≥ 46 g/day than non-drinkers, while for ＊2 allele carriers, CEC levels tended to be lower with a drinking habit of 23-45.9 g/day compared to no drinking habit. Our results suggest that heavy drinking habits may tend to decrease CEC levels, and in the ALDH2 ＊2 allele carriers, even moderate drinking habits may tend to decrease CEC levels.

Association of methylenetetrahydrofolate reductase polymorphism and male infertility in type 2 diabetes mellitus patients

INTRODUCTION: As per World Health Organization, Infertility is a condition which is characterized by not getting conceived with more than 1-year of unprotected sexual intercourse without usage of any contraceptive aids. Folate has a significant role in the metabolism of the cells, like nucleic acids synthesis, gene expression by means of remethylation of the homocysteine into methionine. In the males, deficiency of folate leads to reduced proliferation of the sperm cells. Polymorphisms of methylenetetrahydrofolate reductase (MTHFR) and methionine synthase reductase genes result in alterations of the methylations resulting pathological conditions being a potential risk factor for male infertility. MATERIALS AND METHODS: A total of 100 diabetic infertile males were selected as the study group and 100 nondiabetic fertile males were included as the control group. Blood samples were analyzed for the MTHFR polymorphisms. DNA extraction was done and the extract is subjected to polymerase chain reaction amplification. The resultant is subjected to electrophoresis for MTHFR gene allele confirmation. Statistical analysis was done using IBM SPSS Statistics 20 package. Chi-square test, odds ratio has been done and P &lt; 0.05 is considered statistically significant. RESULTS: Genotype frequencies of MTHFR C677T, A1298C in all the subjects were analyzed for the Hardy–Weinberg law of genetic equilibrium. Mutant polymorphisms of MTHFR C677T, i.e., thymine-thymine and cytosine-thymine were increased significantly. Similarly, MTHFR A1298C mutant polymorphisms, i.e. adenine-cytosine, cytosine-cytosine were increased significantly in the infertile group than in the fertile group. DISCUSSION: Mutations of both MTHFR C677T, A1298C are found to be linked with lowered enzyme activity. We found that the occurrence of mutant homozygous and mutant heterozygous genotypes was increased in a diabetic male infertile group when compared to nondiabetic fertile group. Hence, it can be concluded that presence of single-nucleotide polymorphisms of MTHFR will increase the risk of infertility in diabetic male population.

THE ROLE OF DETERMINANT GENES OF STEROID HORMONES DYSFUNCTION REGULATORS (CYP17A1-RS743572, CYP19A1-RS247015) IN THE DEVELOPMENT OF PCOS IN WOMEN WITH INFERTILITY

Summary. Purpose: to develop prognostic criteria for the outcomes of ART programs in women with infertility in PCOS based on molecular genetic predictors of folliculogenesis disorders. Material and research methods. To solve the tasks set in the work, 125 women were examined: group first - 45 women with primary PCOS and infertility; group second - 46 women with infertility and PCOS in preparation for ART; group third 26 conditionally healthy women. Based on the foregoing, we presented the data of our own studies on the assessment of the state, the genes of steroid hormones (CYP17A1-rs743572, CYP19A1-rs247015) based on the analysis of laboratory data.&#x0D; Results and its discussion. Regarding the influence of the polymorphism of the CY19A1 rs2470152 gene on the development of PCOS, mutant alleles were found to be significantly higher in patients than in the control group. When we divided the main group into MC(+) and MC(-) in terms of CYP19A1 and compared with the control group, we found that the homozygous mutant genotype was found to be greater in the MC(+) and MC(-) group compared to the control group. With this, we can conclude that the homozygous mutational form of the CYP19A1 gene plays a convincing inducible role in PCOS and our result was significant (chi2 - 5.74; p&lt;0.02 in the main group; chi2=5.2; p=0.02 in patients with metabolic syndrome and chi2=3.9; p&lt;0.05 in patients without metabolic syndrome). However, the study did not reveal an induced effect on the heterozygous genotype in the development of PCOS (chi2&lt;3.85; p&gt;0.05). At the same time, the wild-type homozygous variant played a protective role in terms of the appearance of PCOS in the main group, as well as in the MC (+) and MC (-) groups (OR≥0.5). When it comes to the Hardy Weinberg equation, we found no significant difference between the expected and observed results in the main group. Estimates of polymorphism prediction efficiency, as already mentioned, showed only 0.6, which means that the prediction efficiency was not reliable in terms of mutant allele and genotype.&#x0D; In conclusion, from the study of CY17A1 polymorphism in patients with PCOS, it can be said that the GG mutant genotype was statistically significantly more common in patients compared to the control group. When dividing PCOS patients, they were divided into groups and compared with the control group, MS+ PCOS patients had a lower level of the mutant form (GG) genotype compared to the control group, but in MS-PCOS patients compared to the control group, the mutant gene was determined more.

Association of vitamin D deficiency and vitamin D receptor (VDR) gene single-nucleotide polymorphism (rs7975232) with risk of preeclampsia

Background Preeclampsia has a multifactorial-yet-elusive etiology. Recent reports suggest a link between preeclampsia and vitamin D (VD) metabolic axis. Genetic variations like single-nucleotide polymorphisms (SNPs) of vitamin D receptor (VDR) gene can alter the metabolic role of VD, which have been shown by several genetic association studies. However, there is discordance among these studies. Objective The current study aimed to investigate the association of VDR gene polymorphism (ApaI) and VD deficiency with risk of developing preeclampsia. Patients and Method In this case–control study, 40 preeclamptic and 40 normotensive pregnant women were compared for VD status and VDR gene polymorphism. Serum 25-hydroxyvitamin-D [25(OH) D] level was determined by enzyme-linked immunosorbent assay (ELISA) and VDR gene polymorphism Apa1 was analyzed by Allele specific polymerase chain reaction (AS-PCR) using sequence specific primers. Results Serum levels of 25(OH) D were very low but comparable in both preeclamptic and normotensive pregnant women. The difference between the two groups were not statistically significant (p = .423). VDR gene polymorphism ApaI (rs7975232) was found not to have significant association with the risk of developing preeclampsia. The frequencies of wild genotype (GG) in preeclamptic and normotensive women were 27.5% and 22.5% respectively. A total of 25% of preeclamptic women had mutant homozygous genotype (TT) and 17.5% of normotensive women had mutant homozygous genotype. The frequency of mutant heterozygous genotype (GT) in preeclamptic patients was 47.5% and in normotensive women was 60%. The variation of wild and mutant genotypes between the two groups was not statistically significant (p > .05). Conclusion This study showed that VDR gene polymorphism (ApaI) and VD deficiency are not associated with the risk of preeclampsia.

Association of nonsynonymous SNPs of nucleotide excision repair genes ERCC4 rs1800067 (G/A) and ERCC5 rs17655 (G/C) as predisposing risk factors for gallbladder cancer.

Deregulation of DNA repair mechanisms have been frequently demonstrated in the pathology of cancers including gallbladder cancer. We aimed to investigate the association of ERCC4 rs1800067 (G/A) and ERCC5 rs17655 (G/C) with the predisposition in gallbladder cancer and its prognosis. We have also investigated the prognostic and diagnostic values of expression profiles of ERCC4 and ERCC5 in GBC. Polymorphisms of rs1800067 and rs17655 were genotyped by PCR-RFLP. The expression of these genes was analyzed by semi-quantitative PCR. Overall survival was analyzed using Kaplan-Meier plot and cox-regression analysis. Patients with risk group genotypes of rs17655 have shorter overall survival in patients with presence of gallstone, T1+T2 tumor invasion, absence of lymph node involvement and early stages of tumor. Homozygous wild genotype (GG) of rs1800067 and homozygous mutant genotype (CC) of rs17655 together increases two-fold risk of the disease. The variant genotypes (GC/CC) of rs17655 show significantly higher level of ERCC5 expression. Major allele of ERCC4 rs1800067 and minor allele of ERCC5 rs17655 are significantly associated with increased risk of GBC. Upregulation of ERCC4 and ERCC5 is an early event of development of GBC.

Study the relationship between polymorphisms of pre-miRNA regions and expression of mature miRNA in samples of Iraqi infertile men

Polymorphisms that increase risk factor for male infertility is one of the most studied infertility genetic factor however polymorphisms within pre- miRNA sequences have not extensively documented. This study aimed to highlighted association between idiopathic male infertility and, hsa-mir-196a-2 rs11614913Polymorphism genotyping and expression level of mature miRNA were done in semen samples of Iraqi fertile and infertile male in order to study impact of polymorphism on expression level of mature miRNA andstudy correlationbetween both of polymorphism genotype and mature miRNA expression level with quality of semen parameters. Results of this study demonstrated that homozygous mutant (TT) and heterozygous mutant (CT) genotypes of pre-miRNA regions (hsa-miRNA-196a-2, rs11614913) were significantly (P ≤ 0.01) higher in patients group when compared with control and associated with idiopathic Asthenozoospermia, Oligoasthenozoospermia and specifically Azoospermia.Correlation between polymorphism genotype and semen parameters exhibits that genotype obviously impacts semen parameters. Analyzing correlation between polymorphism genotype and expression elucidated that mutant homozygous (TT) genotype increasing expression level of mature miRNA. Elevated level of miRNA-196a-5p exhibitednegative correlation with semen parameter.

Association of SRXN1 Receptor Gene Polymorphism with Susceptibility to Periodontitis.

Emerging evidence suggests that oxidative stress forms a key component in the etiopathogenesis of periodontitis. Literature evidence have shown potential antioxidants responsible for combating the pro-oxidants which stress the periodontium, but the peroxiredoxin-sulfiredoxin system is explored very minimally in periodontal disease. Thus, the present study was aimed to evaluate the genetic association of SRXN1 receptor gene polymorphism (rs6053666). A total of 100 subjects were recruited for this study, which included 50 Periodontitis patients (Stage II and above based on the criteria of American Association of Periodontology-2018) and 50 periodontally healthy or mild gingivitis. Genomic DNA was extracted from the whole blood collected from the subjects. DNA was amplified using specific primers flanking the BtgI region of the SRXN1 receptor gene. The amplicon was further subjected to genotyping using restriction fragment length using BtgI enzyme. The genotype obtained based on the restriction fragment length polymorphism pattern was recorded and used for statistical analysis. The distribution of genotypes and allele frequencies in the periodontitis and control groups were compared using the Chi-square test. The risk associated with individual alleles or genotypes was calculated as the odds ratio with 95% confidence intervals. Statistical significance in all tests was determined at P < 0.05. The genotype frequency and distributions of SRXN1 receptor BtgI polymorphism did not differ significantly at ꭕ2df (P = 0.557). Our study results showed that homozygous and heterozygous mutant genotypes had no significant difference (CC vs. CT + TT) between the periodontitis patients and control group with a P = 0.4266. The detected frequency of CT (38% vs. 34%) and TT (42% vs. 52%) genotype showed no significant difference between control and test group. There was no significant difference in C allele (39% vs. 31%) and T allele (61% vs. 69%) between the test and control group. The present study denotes that SRXN1 receptor gene polymorphism is not associated with periodontitis in the study group analyzed.