ARMS-PCR Method Research Articles

Investigating the Association of rs4962416 and rs6465657 with Prostate Adenocarcinoma in the Iranian Population

Background: Prostate cancer is one of the most commonly diagnosed malignancies in the developed world. Despite other risk factors like age, diet, environment and the pathogenesis of prostate cancer, recent advances in molecular genetics suggest that genetic inheritance plays an important role in prostate cancer. Objective: We attempted to analyze the association of SNPs rs4962416 and rs6465657 in the development of prostate cancer. A better understanding of the association of SNPs in prostate cancer susceptibility may improve risk prediction, improve precision mapping, and provide new insights into the underlying pathophysiology of prostate cancer. To date, no one has investigated these two SNPs in the Iranian populations, and according to the heterogeneity that exists, SNPs in communities should be examined separately. Methods: This case-control study includes 82 people with prostate adenocarcinoma as cases and 96 people with benign prostatic hyperplasia (BPH) as controls. Genotyping of each participant was done by TETRA ARMS-PCR method and for statistical analysis chi-squared, Fisher’s exact logistic regression was used to find the SNPs associated with prostate cancer. Result: The frequency of the polymorphisms rs4962416 and rs6465657 in the prostate adenocarcinoma group was evaluated compared to the BPH control group (p-value < 0.05%) to choose the meaningful SNP. For rs4962416, we didn’t find any meaningful association with prostatic cancer (p=0.402) but for rs6465657 there was a significant difference between genotype frequency (p=0.001). Conclusion: rs6465657 polymorphism which is associated with prostate cancer, can be chosen as a biomarker for this cancer and there should be more investigation on this SNP as these results need to be confirmed in a larger population.

Evaluation of an identification method for the SARS-CoV-2 Delta variant based on the amplification-refractory mutation system.

The Delta variant of SARS-CoV-2 dominated the COVID-19 pandemic due to its high viral replication capacity and immune evasion, causing massive outbreaks of cases, hospitalizations, and deaths. Currently, variant identification is performed mainly by sequencing. However, the high requirements for equipment and operators as well as its high cost have limited its application in underdeveloped regions. To achieve an economical and rapid method of variant identification suitable for undeveloped areas, we applied an amplification-refractory mutation system (ARMS) based on PCR for the detection of novel coronavirus variants. The results showed that this method could be finished in 90min and detect as few as 500 copies/mL and not react with SARS-Coronavirus, influenza A H1N1(2009), and other cross-pathogens or be influenced by fresh human blood, α- interferon, and other interfering substances. In a set of double-blind trials, tests of 262 samples obtained from patients confirmed with Delta variant infection revealed that our method was able to accurately identify the Delta variant with high sensitivity and specificity. In conclusion, the ARMS-PCR method applied in Delta variant identification is rapid, sensitive, specific, economical, and suitable for undeveloped areas. In our future study, ARMS-PCR will be further applied in the identification of other variants, such as Omicron.

Biochemical association of regulatory variant of KLF14 genotype in the pathogenesis of cardiodiabetic patients.

The study focuses on examining the relationship between a single nucleotide polymorphism (SNP) in KLF14 rs4731702 and risk of type 2 diabetes mellitus (T2DM) and dyslipidemia in different ethnic populations. The purpose of this study was to evaluate the association between KLF14 rs4731702 and serum lipid profile and to determine the frequency distribution of KLF14 rs4731702 among T2DM and cardiometabolic patients. A total of 300 volunteers were recruited, consisting of three groups: 100 healthy individuals, 100 individuals diagnosed with T2DM, and 100 individuals diagnosed with cardiometabolic disorders. Biochemical analysis of blood samples was conducted to assess various biomarkers related to glycemic control and lipid profile. This involved measuring levels of glucose, triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and ApoA1. Genotyping analysis was performed to investigate KLF14 rs4731702 polymorphism. The Tetra ARMS-PCR method was employed for genotyping analysis. The results of biochemical profiling revealed a significant association between altered glycemic biomarkers and lipid profile in diseased patients compared to healthy participants. The frequencies of KLF14 rs4731702 alleles and genotypes were compared between the control group and T2DM group. A statistically significant difference was observed, indicating a potential association between KLF14 rs4731702 and T2DM. In the dominant inheritance model of KLF14 rs4731702 SNP, a statistically significant difference [odds ratio (95% confidence interval)] of 0.56 (0.34 -0.96) was found between the control and T2DM subjects. This suggests that the presence of certain genotypes influences the risk of T2DM. In T2DM patients, individuals carrying the C allele exhibited compromised insulin sensitivity, decreased HDL-C and ApoA1 levels, and increased serum glucose, TG, and LDL-C concentrations. Conversely, TT genotype carriers demonstrated increased levels of HDL-C and ApoA1, lower insulin resistance, serum glucose, LDL-C, and TG levels. The study's findings indicate that dyslipidemia in T2DM patients is associated with reduced KLF14 functionality due to CC and CT genotypes, leading to insulin resistance and an increased risk of cardiovascular diseases. Additionally, risk of KLF14 rs4731702 polymorphism was found to increase with age and was more prevalent in female than in male individuals. These insights contribute to understanding genetic factors influencing the development and progression of T2DM and dyslipidemia in different ethnic populations.

The rs9402373 polymorphism of CTGF gene may not be related to inflammatory bowel disease susceptibility in Chinese population based on ARMS-PCR genotyping

BackgroundIt has been confirmed that the connective tissue growth factor (CTGF) gene rs9402373 polymorphism is associated with fibrotic and inflammatory diseases. However, studies on the relationship between polymorphisms in CTGF rs9402373 and inflammatory bowel disease (IBD) remain rare. Therefore, the aim of this study was to assess the association between the CTGF rs9402373 polymorphism and IBD susceptibility in a Chinese population. Materials and methodsTo establish an amplification refractory mutation system (ARMS) PCR technology for genotyping CTGF gene rs9402373 polymorphism, we designed two specific forward primers for the wild and mutant types by placing the allele-specific nucleotide at the penultimate position of the ‘3′ end of the primer. Then, 10 samples were randomly selected and rechecked by DNA sequencing to verify the accuracy of this method. We further used the established method to detect specimens collected from 191 patients with inflammatory bowel disease, including 120 Crohn's disease (CD) and 71 ulcerative colitis (UC), and 110 healthy Han Chinese individuals. ResultsWe successfully established the ARMS-PCR method for genotyping, and the results of 10 randomly selected samples were completely consistent with DNA sequencing. The rs9402373 G allele frequencies in UC and CD cases were 38.03% and 43.75%, respectively, and in controls, they were 41.82%. No significant difference was found in minor allele frequencies between the UC or CD and control groups (P = 0.473, P = 0.676). Genotype analysis demonstrated that there was no relationship between CTGF rs9402373 polymorphism and the risk of IBD regardless of the inheritance mode (P > 0.05). ConclusionsIn this preliminary study, we successfully developed a simple, efficient and cost-effective method for genotyping CTGF rs9402373 polymorphism. The polymorphism may not be related to IBD susceptibility in the Chinese Han population.

Analysis of KLF7 and KLF5 transcription factors gene variants in coronary artery disease

Introduction and objectiveGenetic susceptibility has a key role in the pathogenesis of coronary artery disease (CAD). KLF5 and KLF7 are transcriptional factors essential to cell development and differentiation. Their genetic variants have been associated with the risk of metabolic disorders. The present study aimed to evaluate the possible correlation of KLF5 (rs3812852) and KLF7 (rs2302870) single nucleotide polymorphisms (SNPs) with the risk of CAD for the first time in the world. MethodsThe clinical trial study comprised 150 patients with CAD and 150 control subjects without CAD from the Iranian population. After blood sampling, deoxyribonucleic acid was extracted and genotyped using the Tetra Primer ARMS-PCR method and confirmed by Sanger sequencing. ResultsThe KLF7 A/C genotypes and C allele frequency were meaningfully higher in the control group compared to the CAD+ group (p<0.05). No obvious association has been observed between KLF5 variants and CAD risk. However, the distribution of the AG genotype of KLF5 was statistically lower in CAD+ patients with diabetes than in CAD+ patients without diabetes (p<0.05). ConclusionThis study identified KLF7 SNP as a causative gene contributing to CAD, which presents novel insight into the molecular pathogenesis of the disease. It is, however, unlikely that KLF5 SNP has an essential role in the risk of CAD in the studied population.

Genetic analysis links the diagnosis of carriers of the PKD1 mutation gene that causes polycystic kidney disease

Polycystic kidney disease (PKD) is an inherited disorder characterised by the formation and growth of multiple cysts in the kidney, accompanied by a gradual increase in the size of both kidneys leading to impaired kidney function and renal failure. Autosomal dominant PKD is the most common genetic disorder with an estimated prevalence of 1/500-1/1000 in Western countries [1], caused by mutations in the PKD1 (MIM#601313) and PKD2 (MIM#173910), of which 85% of cases are caused by mutations in the PKD1 gene [2]. Because of the PKD1gene’s enormous size, numerous mutations, and up to 6 structural pseudogenes, which share a high degree of similarity sequences (97.7%) from the 5’UTR to exon 32 [3, 4], the methods for identifying PKD1 mutations are currently restricted and challenging from amplifying the mutant gene area. Therefore, it is essential to create a novel diagnostic technique with high accuracy and broad applicability for various mutation types that may circumvent the shortcomings of the conventional method. Objective: To develop a genetic linkage analysis technique for detecting the person with PKD1 mutation causing PKD. Materials and methods: 11 members were from a family with PKD1 gene mutations causing PKD. Designed PKD1 gene-linked STRs primers and conducted genetic linkage analysis with the family members. After comparing the outcomes of the technique with those of the Touchdown-PCR and ARMS-PCR methods, the complete technique is subsequently given. Results: Successfully developed the genetic linkage analysis technique for detecting the person with PKD1mutation causing PKD.

MTHFR and MTRR Genetic Polymorphism of Methotrexate Therapy Outcomes in Early Rheumatoid Arthritis.

Methotrexate (MTX) is used as an anchor drug for the treatment of rheumatoid arthritis (RA) and there may be differences in drug action between genotypes. The purpose of this study was to investigate the relationship between clinical efficacy response and disease activity of MTX monotherapy with methylenetetrahydrofolate reductase (MTHFR) and methionine synthase reductase (MTRR) polymorphisms. In the study, a population of 32 patients in East China with early RA fulfilling the diagnostic standards of the American College of Rheumatology (ACR) were enrolled, all of them received MTX monotherapy. Genotyping of patients MTHFR C677T and A1298C, MTRR A66G using tetra-primer ARMS-PCR method and sanger sequencing to verify its accuracy. The distribution of three polymorphic genotypes that were studied is in accordance with the Hardy-Weinberg genetic equilibrium. The patient pathology variables smoke (OR = 0.088, P = 0.037), drink alcohol (OR = 0.039, P = 0.016) and males (OR = 0.088, P = 0.037) were significantly associated with non-response to MTX. Genotype, allele distribution and genetic statistical models were not found to be related to MTX treatment response and disease activity in both the response groups and non-response groups. Our findings suggest that the MTHFR C677T, MTHFR A1298C and MTRR A66G polymorphisms may not predict MTX clinical treatment response and disease activity in patients with early RA. The study revealed that smoke, alcohol, and males were possible influential factors for MTX non-response.

The association of paraoxonase 1 activities, serum mRNA expression and polymorphisms with in-stent coronary restenosis; a case-control study

BackgroundPON1 polymorphisms and enzyme activity have been reported to be associated with cardiovascular disease (CVD). The aim of current study was to assess the association of these PON1 SNPs and serum enzyme activity and with restenosis following coronary artery stenting. MethodsA total of 306 unrelated Iranian patients with history of coronary stent implantation who underwent elective re-angiography were enrolled in the current study. Three single nucleotide polymorphisms (SNPs) of PON1 gene were genotyped. We used a double ARMS PCR method for genotyping at the Q192R and L55M loci, and PCR-RFLP method for C-108 T. Serum mRNA expression of PON1 was determined using SYBER green method and Roche LightCycler. ResultsThere were 104 patients considered as in-stent restenosis (ISR) (mean age: 60.55 ± 8.60 years), and 202 individuals as non ISR (NISR) (mean age: 61.94 ± 9.18 years). There were no significant difference between paraoxonase and arylestrase actvities of paraoxonase 1 enzyme. Furthermore, there were no significant differences in the distribution of genotypes of PON1 polymorphism with ISR compared to the control group. The AAC haplotype was the most frequent haplotype in our population study, and haplotypes were not associated with ISR occurrence. The difference between serum mRNA expression of PON1 was not significant between ISR and NISR groups. ConclusionsAccording to our finding it seems that paraoxonase 1 activities, the most important polymorphisms of PON1 including Q192R, L55M and C-108 T and serum mRNA expression of PON1 are not beneficial markers for ISR diagnosis.

Assessment of the Potential Role of Selected Single Nucleotide Polymorphisms (SNPs) of Genes Related to the Functioning of Regulatory T Cells in the Pathogenesis of Psoriasis

Recent studies have indicated a key role of the impaired suppressive capacity of regulatory T cells (Tregs) in psoriasis (PsO) pathogenesis. However, the genetic background of Treg dysfunctions remains unknown. The aim of this study was to evaluate the association of PsO development with selected single nucleotide polymorphisms (SNPs) of genes in which protein products play a significant role in the regulation of differentiation and function of Tregs. There were three study groups in our research and each consisted of different unrelated patients and controls: 192 PsO patients and 5605 healthy volunteers in the microarray genotyping group, 150 PsO patients and 173 controls in the ARMS-PCR method group, and 6 PsO patients and 6 healthy volunteers in the expression analysis group. The DNA microarrays analysis (283 SNPs of 57 genes) and ARMS-PCR method (8 SNPs in 7 genes) were used to determine the frequency of occurrence of SNPs in selected genes. The mRNA expression of selected genes was determined in skin samples. There were statistically significant differences in the allele frequencies of four SNPs in three genes (TNF, IL12RB2, and IL12B) between early-onset PsO patients and controls. The lowest p-value was observed for rs3093662 (TNF), and the G allele carriers had a 2.73 times higher risk of developing early-onset PsO. Moreover, the study revealed significant differences in the frequency of SNPs and their influence on PsO development between early- and late-onset PsO. Based on the ARMS-PCR method, the association between some polymorphisms of four genes (IL4, IL10, TGFB1, and STAT3) and the risk of developing PsO was noticed. Psoriatic lesions were characterized with a lower mRNA expression of FOXP3, CTLA4, and IL2, and a higher expression of TNF and IL1A in comparison with unaffected skin. In conclusion, the genetic background associated with properly functioning Tregs seems to play a significant role in PsO pathogenesis and could have diagnostic value.

Human papillomavirus infection and its relationship with common polymorphism of HLA gene by PCR method

Studies have shown that human papillomavirus (HPV) causes cervical cancer, but investigations on the relationship between HPV and HLA gene polymorphism are limited.Objective: The purpose of this study is to investigate the prevalence of HPV infection and its relationship with common HLA gene polymorphisms in cervical cancer patients using the PCR method. This study used 52 fixed samples from the laboratory of Khatam al-Anbia Hospital in Tehran. DNA was isolated using proteinase K and a DNPTM commercial extraction kit. The PCR technique was used to identify L1, HPV-16, HPV-18, HPV-31, HPV-11, HPV-33, and HPV-35 genes. The ARMS-PCR method was used to determine the genotypes of common HLA polymorphisms. The data were analyzed using one-way analysis of variance. The ARMS-PCR results showed that 10 polymorphisms were observed in the HLA gene in the samples, with 7 related to DRB1, 2 related to DQB1, and 1 related to DQA1. However, the desired polymorphisms were not observed in 42 of the studied samples. Polymorphism of DRB1 genes in 7 samples showed that 6 of these 7 samples were infected with HPV16 and one with HPV18. The study results suggest that HPV16 and HPV18 are the most common HPV types isolated from cervical cancer patients. HLA gene polymorphism may predispose people to these infections. Understanding these relationships in different populations and races is essential for understanding the role of HLA gene polymorphism in cervical cancer and its relationship with HPV infection.

TMPRSS2 polymorphism (rs12329760) and the severity of the COVID-19 in Iranian population.

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has been responsible for the recent pandemic since early 2020. Due to the wide range of clinical symptoms of this disease, from asymptomatic to severe and critical forms, it seems that genetic differences among patients, along with other factors (such as gender, age, and underlying diseases), can explain part of the variation in disease symptoms. The TMPRSS2 enzyme plays a vital role in the early stages of the interaction of the SARS-CoV-2 with the host cells by facilitating viral entry. There is a polymorphism in the TMPRSS2 gene, called rs12329760(C to T) as a missense variant, which causes the replacement of valine to methionine in the TMPRSS2 protein at position 160. The present study investigated the association between the TMPRSS2 genotype and the severity of the Coronavirus disease 2019 (COVID-19) in Iranian patients. The TMPRSS2 genotype of 251 COVID-19 patients (151 patients with asymptomatic to mild and 100 patients with severe to critical symptoms) was detected on genomic DNA extracted from patients' peripheral blood via the ARMS-PCR method. Our results showed a significant association between the minor T allele and the severity of the COVID-19 (P-value = 0.043) under the dominant and additive inheritance model. In conclusion, the results of this study showed that the T allele of the rs12329760 in the TMPRSS2 gene is a risk allele for severe form of COVID-19 in Iranian patients in contrast to most previous studies on this variant in European ancestry populations which suggested this variant as a protective allele. Our results reiterate to the ethnic-specific risk alleles and hidden unknown complexity behind the host genetic susceptibility. However, further studies are needed to address the complex mechanisms behind the interaction of the TMPRSS2 protein and the SARS-CoV-2 and the role of rs12329760 polymorphism in determining the disease severity.

Association study to evaluate Foxo1 and Foxo3 gene polymorphisms in polycystic ovary syndrome: a preliminary case-control study and in silico analysis.

Polycystic ovary syndrome (PCOS) is known as a multifactorial and multi-gene-mediated endocrine disorder among women of reproductive age. FoxO1 and FoxO3 are members of the forkhead transcriptional factors family that play a pivotal role in the function of ovaries. The current work is aimed at investigating the association between gene variants of FoxO1 and FoxO3 and the risk of PCOS in a sample of the Iranian population. We recruited 200 women diagnosed with PCOS and 200 healthy women. Both polymerase PCR-RFLP and ARMS-PCR methods were used for genotyping. Sanger sequencing was recruited to confirm the genotypingresults. The T allele of rs17592236 and the C allele of rs12585277 decreased PCOS risk by 29 and 28%, respectively. In contrast, the C allele of rs2253310 and G allele of rs2802292 increasedthe risk of PCOS by 1.39 and 1.63 folds, correspondingly. Bioinformatics results showed that some genes, including matrix metallopeptidase 9 (MMP-9), phosphoinositide-3-Kinase Regulatory Subunit 224 1 (PIK3R1), peroxisome proliferator-activated receptor Gamma (PPARG), and glycogen synthase 225 kinase-3 beta (GSK-3 beta) have significant interactions with FoxO1, suggesting that FoxO1 might have crucial roles in regulating different signaling pathways in ovarian cells. We found that FoxO1 rs17592236C > T and rs12585277C > T had a protective role against PCOS, while FoxO3 rs2253310C > G and rs2802292G > T enhanced the risk of this metabolic disorder in our population. Additional studies on larger populations with varying racesare needed to confirm these findings.

Spectrum of Rare and Novel Indel Mutations Responsible for β Thalassemia in Eastern India.

The online version contains supplementary material available at 10.1007/s12291-022-01098-w.

Genetic variation in C-reactive protein (CRP) gene is associated with retinopathy and hypertension in adolescents with type 1 diabetes.

Elevated concentration of CRP has been associated with the risk of diabetes as well as cardiovascular events and microvascular complications in T1D patients. We hypothesize that the +1846 C&gt;T CRP gene polymorphism may have impact on the risk of T1D and/or its complications. We have examined 400 young patients with T1D and 250 healthy age-matched controls. The +1846 C&gt;T CRP gene polymorphism was genotyped by ARMS-PCR method. The analysis covers microvascular complications, concentrations of serum pro- and anti-inflammatory markers, adhesion molecules, proangiogenic factor as well as blood pressure. CT genotype (OR=1.799) and T allele (OR=1.733) are associated with increased risk of T1D, while CC genotype decreases the risk of this condition (OR=0.458). Moreover, increased risk of hypertension corresponds with TT and T variant (OR=3.116 and OR=1.830, resp.) while CC genotype is decreasing the risk (OR=0.547). Furthermore, CT variant is connected with lower risk of retinopathy (OR=0.512) whereas TT variant decreases the risk of this complication (OR=2.228). Our data also implies various effects of CRP +1846 C&gt;T polymorphism on the inflammatory status of T1D patients. Although further studies are required, the +1846 C&gt;T CRP gene polymorphism could be considered a genetic marker to predict susceptibility to retinopathy and hypertension in T1D adolescents.

The Effect of Caspase 8, 9 Gene Polymorphisms on Non-Hodgkin Lymphoma Susceptibility and Clinical/Pathological Features

Caspases (CASPs) are the main executors of the apoptotic process. Studies to date have shown the role of caspase-8 (CASP8) and caspase-9 (CASP9) in carcinogenesis. Therefore, the aim of this study was to investigate the associations between CASP9-rs4233532, CASP9-rs4646018, and CASP8- rs1045485 gene polymorphisms and non-Hodgkin lymphoma (NHL) susceptibility in an Iranian population-based study. Moreover, it was examined whether such the genotype of these polymorphisms is related with clinicopathological characteristics of NHL. 175 patients with NHL and 175 age- and sex-matched controls were enrolled in this study. We determined the genotypes of single nucleotide polymorphisms (SNPs) from CASP genes with Tetra ARMS-PCR (Amplification refractory mutation system) method. Statistically significant association were observed between CASP9-rs4646018 and increased risk of NHL under codominant CC, codominant TC, and dominant TC+CC genetic models. Our results showed that the A allele of CASP8-rs1045485 was a protective factor for NHL and GArs1045485 genotype significantly reduced risk of NHL. In contrast, CASP9- rs4233532 was not linked to NHL susceptibility. No relationship was detected between CASP8-rs1045485 and CASP9-rs4233532 and NHL clinicopathological characteristics, however genetic variation in CASP9-rs4646018 was associated with histology, treatment and radio therapy of NHL. Our study presented that the CASP8- rs1045485 and CASP9-rs4646018 polymorphisms could affect the risk of NHL in Iranian populations which was the first report to show the significant relationship between rs1045485, rs4646018 polymorphisms and NHL susceptibility. Replication large-scale case-control studies in different ethnicities are warranted to verify these findings.

Coding Variants of the FMO3 Gene Are Associated with the Risk of Chronic Kidney Disease: A Case-Control Study.

Chronic kidney disease (CKD) is a global health concern involving roughly one-tenth of developed countries' populations. The flavin-containing dimethylaniline monooxygenase 3 (FMO3) gene encodes an enzyme that catalyzes trimethylamine N-oxide (TMAO), a toxin in CKD sufferers. This preliminary study aims to evaluate the association between coding region variations of FMO3, rs2266782G/A (E158K), rs2266780A/G (E308G), and rs1736557G/A (V257M), and the susceptibility to CKD. A total of 356 participants were enrolled, including 157 patients diagnosed with CKD and 199 age-matched healthy individuals. Genotyping of FMO3 gene variations was performed via PCR-RFLP and ARMS-PCR methods. Our findings revealed a significant association between rs2266780A/G and rs1736557G/A and CKD under different genetic models. Compared to the GGG haplotype of rs2266782/rs1736557/rs2266780, the GAG, GAA, AAG, and AAA haplotype combinations conferred an increased risk of CKD in our population. Interaction analysis revealed that some genotype combinations, including GA/AA/AA, AA/AA/AA, GA/AA/GA, and GG/AG/AA, dramatically increased CKD risk in the Iranian population. No correlation was found between FMO3 polymorphisms and CKD stages. These observations highlight the potential impact of coding variants of the FMO3 gene on the onset of CKD. Further investigations into expanded populations and diverse races are needed to confirm our findings.

Association of CXCL12 and CXCR4 Genes Polymorphism with Proliferative Diabetic Retinopathy in Diabetic Patients

Introduction: Patients with diabetes are prone to develop complications related to the retina. So far, several genes have been reported in connection with the progression of retinopathy, and a group of these genes is genes involved in angiogenesis. CXCL12 and CXCR4 genes are among the most important genes in this path, which have been less studied and investigated. The aim of this study was to determine the relationship between polymorphism of CXCL12 (rs1801157) and CXCR4 (rs2228014) genes and the risk of proliferative diabetic retinopathy in the population of Yazd City.&#x0D; Methods: In the present case-control study, rs2228014 and rs1801157 polymorphisms were investigated among 103 patients with diabetes in two groups, case and control. In this study, the control group included 49 diabetic patients who did not show retinopathy complications, and the case group consisted of 54 diabetic patients with proliferative retinopathy complications. The Genotyping of rs2228014 polymorphism was examined by ARMS-PCR method and rs1801157 variant by RFLP-PCR technique. The statistical analysis of data was conducted using SPSS software version 16 and chi-square test.&#x0D; Results: Our results represented no association among the rs1801157 and rs2228014 polymorphisms and the risk of proliferative diabetic retinopathy (P&gt;0.05).&#x0D; Conclusion: This study did not confirm association of the rs1801157 and rs2228014 polymorphisms with proliferative diabetic retinopathy and further studies are required to investigate the correlation of polymorphisms of interest with PDR.

Genetic Polymorphisms in miR-137 and Its Target Genes, TCF4 and CACNA1C, Contribute to the Risk of Bipolar Disorder: A Preliminary Case-Control Study and Bioinformatics Analysis.

Accumulating evidence has suggested that miR-137 and its target genes, CACNA1C, and TCF4, are amongst the most robustly implicated genes in psychiatric disorders. This preliminary study is aimed at investigating the effects of genetic variations in miR-137 (rs1625579A/C), TCF4 (rs1261084C/T), and CACNA1C (rs10774053A/G and rs10466907G/T) on BD susceptibility. We recruited 252 BD patients and 213 healthy subjects as the control group. Genotyping was performed using PCR-RFLP and ARMS-PCR methods. Enhanced risk of BD was found under the codominant homozygous, dominant, and allelic models of TCF4 rs1261084C/T, codominant homozygous and allelic models of CACNA1C rs10466907G/T polymorphisms, as well as codominant homozygous, dominant, recessive, and allelic models of the CACNA1C rs10774053A/G. Moreover, both TT/AG/GT/AA and TT/GG/GT/AC genotype combinations strongly increased the risk of BD in the participants. The bioinformatics analyses revealed that rs1261084C/T and rs10466907G/T created and disrupted binding sites of some miRNAs in the 3′-untranslated region of TCF4 and CACNA1C genes. In contrast, the rs10774053A/G created a new binding site for a major splicing factor and might have an effective role in the function of the CACNA1C protein. We have found that all the studied SNPs are positively associated with BD susceptibility. Replicated studies on different ethnicities are required to confirm these findings.

Association of Paraoxonase-1 Genotype and Phenotype with Angiogram Positive Coronary Artery Disease.

ResumoFundamento Tem sido demonstrado que um aumento dos níveis séricos de PON1 é protetor contra vários distúrbios. Foi relatado que vários polimorfismos de nucleotídeo único (SNPs, single nucleotide polymorphisms ) do gene PON1 estão associados a níveis e atividade de proteínas enzimáticas séricas.Objetivos Investigar a associação de SNPs do PON1 e atividade da paraoxonase sérica com a doença arterial coronariana (DAC).Métodos Foram estudados 601 pacientes não relacionados submetidos à angiografia coronária, incluindo aqueles com estenose >50% (N=266) e aqueles com estenose <30% (N=335). Os SNPs rs662 e rs840560 do gene da paraoxonase foram determinados utilizando o método ARMS-PCR e o SNP rs705379 foi genotipado utilizando análise de PCR-RFLP. A atividade da paraoxonase sérica foi medida utilizando paraoxon como substrato. O valor de p<0,05 foi considerado significante.Resultados A atividade da paraoxonase sérica não foi significativamente diferente entre os grupos de estudo. Após ajuste para idade, sexo, hipertensão, diabetes mellitus e dislipidemia, o genótipo GG e o modelo codominante de rs662 foram positivamente associados a uma angiografia positiva (respectivamente, OR = 2,424, IC 95% [1,123-5,233], p <0,05, OR = 1,663, IC 95% [1,086-2,547]). A atividade da paraoxonase sérica foi significativamente maior no alelo G e variante GG do polimorfismo rs662, alelo A e variante AA de rs854560 e alelo C e variante CC de rs705379. A análise de haplótipos mostrou que o haplótipo ATC foi significativamente mais prevalente no grupo com angiografia negativa. A análise entre os grupos indicou que o alelo A de rs662 foi significativamente associado à menor atividade da paraoxonase no grupo com angiografia positiva (p=0,019).Conclusões A presença do alelo G do polimorfismo de nucleotídeo único rs662 está independentemente associada ao aumento do risco de DAC.

The association of APOE genotype with COVID-19 disease severity

COVID-19 has caused the recent pandemic of respiratory infection, which threatened global health. The severity of the symptoms varies among affected individuals, from asymptotic or mild signs to severe or critical illness. Genetic predisposition explains the variation in disease severity among patients who suffer from severe symptoms without any known background risk factors. The present study was performed to show the association between APOE genotype and the severity of COVID-19 disease. The APOE genotype of 201 COVID-19 patients (101 patients with asymptomatic to mild form of the disease as the control group and 100 patients with severe to critical illness without any known background risk factors as the case group) were detected via multiplex tetra-primer ARMS-PCR method. Results showed that the e4 allele increased the risk of the COVID-19 infection severity more than five times and the e4/e4 genotype showed a 17-fold increase in the risk of severe disease. In conclusion, since our study design was based on the exclusion of patients with underlying diseases predisposing to severe form of COVID-19 and diseases related to the APOE gene in the study population, our results showed that the e4 genotype is independently associated with the severity of COVID-19 disease. However, further studies are needed to confirm these findings in other nations and to demonstrate the mechanisms behind the role of these alleles in disease severity.