4G Allele Research Articles

ALLELIC VARIANTS OF MATRIX METALLOPROTEINASE TIPES 1 AND 3 GENES IN PRESCHOOL CHILDREN SUFFERING FROM RECURRENT RESPIRATORY

Matrix metalloproteinases (MMPs) have numerous physiological and pathological effects. In particular, they are involved in extracellular matrix degradation and remodeling, cell differentiation and apoptosis, immune and inflammatory mechanisms, embryogenesis, etc. Aim. To determine the state of interdependence between the allelic variants of the genes MMP-1-1607 (1G/2G) and MMP-3-1171 (5A/6A), on the one hand, and the different frequency of acute respiratory infections (ARI) in preschool children, on the other hand. Materials and methods. A total of 33 children (15 boys and 18 girls), aged 1-6 years, undergoing inpatient treatment for ARI were included in the clinical trial. The number of ARI episodes during the previous year of life was regarded. The distribution of allelic variants was also studied, specifically: 1) 1G/2G of the MMP-1 gene at position 1607 (rs 1799750) and 2) 5A/6A of the MMP-3 gene at position 1171 (rs 35068180). Statistical processing of the obtained digital data was performed using IBM SPSS Statistics 28 licensed software (No. 3922/01/2021). Results. The association between preschool children belonging to the subgroup with more frequent ARI episodes and the 2G/2G genotype for the rs 1799750 polymorphism locus of the MMP-1-1607 gene was demonstrated (OR=7.778; p1=0.008; p2=0.025; CI: 1.561 - 38.756). On the contrary, the 1G/2G genotype was significantly more frequent in patients forming a subgroup with fewer ARI episodes (OR=0.127; p1=0.014; p2=0.039; CI: 0.022 - 0.739). Furthermore, the 1G allele was more frequent in children with fewer ARI episodes, and the 2G allele was more frequent in those with a greater number of such episodes in their history (OR=2.778; p1=0.044; p2=0.080; CI: 1.016 - 7.639). At the same time, there was no association between alleles 5A and 6A and their corresponding genotypes, determined for the rs 35068180 polymorphism locus of the MMP-3 gene at position 1171, and subgroups of children with less frequent and more frequent ARI episodes. Conclusions. The results obtained suggest a significant dependence of the frequency of ARI episodes in preschool children on the studied genotypes (2G/2G and 1G/2G) of the MMP-1 gene (rs 1799750). They should be considered in predicting whether such children are at risk for recurrent course of ARI. The study was conducted within the framework of the initiative research work of the Department of Pediatrics with Childhood Infections of Luhansk State Medical University (Rubizhne, Ukraine) - "Current aspects of the influence of perinatal factors on the development of somatic pathology in children aged 1-14 years" (2017-2021, state registration number 0117U003041). No external funding was received for this study. The study was conducted in accordance with the tenets of the Declaration of Helsinki. The study protocol was approved by the local ethics committees of Bogomolets National Medical University and Luhansk State Medical University. Informed parental consent was obtained for the study in children. No conflict of interests was declared by the authors.

The Impact of Emotional Responses on Female Reproduction: Fibrinolysis in the Spotlight.

Fibrinolytic enzymes modify various substrates required for tissue remodeling, playing a crucial role in mechanisms underlying resilience, reward processing, ovulation, embryo implantation, and placentation. Individuals with low resilience and reduced reward responsiveness, when exposed to chronic stress, are at increased risk of experiencing a range of negative emotions. Chronic anxiety and melancholia are examples of negative emotions associated with hypercortisolism, while fear and atypical depression are characterized by systemic inflammation. Both cortisol and inflammatory cytokines stimulate the production of plasminogen activator inhibitor-1 (PAI-1), a potent fibrinolysis inhibitor. Chronic anxiety, fear, and depression are among the many hypofibrinolytic conditions increasing the risk of oligo-anovulation, miscarriage, fetal growth restriction, and preeclampsia. Although significant, the impact of negative emotions on implantation is not as obvious as on ovulation or placentation. Other hypofibrinolytic conditions that may affect female reproduction through mechanisms dependent or independent of PAI-1 include metabolic disturbances (e.g., due to consumption of highly palatable foods, often used to alleviate negative affect), inflammation, hyperhomocysteinemia, hypothyroidism, hypercortisolism, antiphospholipid antibodies, and the 4G allele of the PAI-1 gene. Benzodiazepines and antidepressants should be used with caution in the first trimester as this combination may cause malformations. Also, selective serotonin reuptake inhibitors have fibrinolytic properties that increase the risk of bleeding after surgical procedures. Psychological interventions, especially group therapy, are effective in the prevention of reproductive disorders. Controlled trials are needed to test the hypothesis that female reproductive health depends on psychological well-being, a balanced diet and physical activity, suppression of inflammation and autoantibodies, and homocysteine and hormonal homeostasis.

Global DNA Methylation Levels Viz-a-Viz Genetic and Biochemical Variations in One Carbon Metabolic Pathway: An Exploratory Study from North India.

Despite the importance of one carbon metabolic pathway (OCMP) in modulating the DNA methylation process, only a few population-based studies have explored their relationship among healthy individuals. This study aimed to understand the variations in global DNA methylation levels with respect to selected genetic (CBS 844ins68, MTRR A66G, MTR A2756G, and MTHFR C677T polymorphisms) and biochemical (folate, vitamin B12, and homocysteine) markers associated with OCMP among healthy North Indian adults. The study has been conducted among 1095 individuals of either sex (69.5% females), aged 30-75 years. A sample of 5 mL of blood was collected from each participant. Homocysteine, folate, and vitamin B12 levels were determined using the chemiluminescence technique. Restriction digestion was performed for genotyping MTRR A66G, MTR A2756G, and MTHFR C677T polymorphisms and allele-specific PCR amplification for CBS 844ins68 polymorphism. Global DNA methylation levels were analyzed using ELISA-based colorimetric technique. Of the selected genetic and biochemical markers, the mutant MTRR A66G allele was positively associated with global DNA methylation levels. Further, advanced age was inversely associated with methylation levels. MTRR 66GG genotype group was hypermethylated than other genotypes in folate replete and vitamin B12 deficient group (a condition prevalent among vegetarians), suggesting that the G allele may be more efficient than the wild-type allele in such conditions. Global DNA methylation levels appeared to be more influenced by genetic than biochemical factors. MTRR 66G allele may have a selective advantage in vitamin B12 deficient conditions. Further research should be undertaken to understand how genetics affects epigenetic processes.

A Nomogram Model Containing Genetic Polymorphisms to Predict Risk of Pulmonary Embolism in Pregnant Women.

Pulmonary embolism (PE), the most serious presentation of venous thromboembolism (VTE), is associated with a high rate of mortality and expense. Clinical studies on pregnant women with PE are scarce. The aim of this study was to analyze the clinical impact of fibrinolytic enzyme activation inhibitor-1 (PAI-1) 4G/5G genetic polymorphisms, methylenetetrahydrofolate reductase (MTHFR) rs1801131 (A1298C) and rs1801133 (C677T) genetic polymorphisms, and establish a predictive model for pregnant women. Between September 2022 and August 2023, 53 pregnant women with PE were enrolled. Using the propensity score matching method, 106 consecutive pregnant women without VTE were 1:2 matched. The relevant patient data were collected, and the susceptibility genes for PE were detected to determine genetic polymorphisms, and PE susceptibility in pregnant women, as well as to develop predictive models. Our study showed that 4G/4G homozygous mutations increased the risk of pregnant PE fourfold (OR = 4.46, 95% CI = 1.59-12.50, P = 0.004), whereas the 4G allele mutation increased the risk twofold (OR = 2.33, 95% CI = 1.35-4.04, P = 0.002). A nomogram was established to predict the risk of pregnant women with PE by four predictive features including PAI-1 genetic polymorphisms, international normalized ratio (INR), antithrombin-III (AT-III) activity, and platelet count (PLT). The area under the curve (AUC) of the nomogram was 0.821 (0.744-0.898). The AUC of the internal validation group was 0.822 (0.674-0.971). Decision curve analysis revealed that the nomogram has a higher net benefit in the following threshold: probability interval of ≥15%. The PAI-1 4G/4G genotype is an independent risk factor for pregnant women with PE; furthermore, the presence of the 4G allele can increase the risk of PE. The study established a nomogram to predict the risk of PE in pregnant women.

Thrombophilic gene variants in patients with coronary artery disease and myocardial infarction

Background and aim: Coronary artery disease (CAD) and myocardial infarction (MI) are cardiovascular diseases that occur due to atherosclerosis (plaque formation) or atherosclerotic obstruction of the coronary arteries. Their genetic basis has been under investigation for a long time, and common variant studies link different genetic loci with these diseases. In this study, we investigated the possible association of coronary artery disease and myocardial infarction with thrombophilic gene variants, including MTHFR C677T and A1298C, Beta fibrinogen -455G/A, Factor XIIIV34L and PAI-1 4G/5G single nucleotide polymorphisms (SNPs).
 Methods: A total of 128 people (64 patients and 64 controls) were included in the study. Genomic DNA was isolated using the EZ1 blood mini kit. The DNA was amplified and PCR was performed using the PyroMark PCR Kit (Qiagen, Germany). Pyrosequencing reaction was completed by processing with PyroMark Q24 instrument.
 Results: We found that the PAI-1 4G/5G polymorphism and the 4G allele were significantly associated with coronary artery disease and myocardial infarction (P= 0.01). Although mutant variants were higher in patients, no statistically significant difference was observed between the patient and control groups in terms of FXIII, Beta-fibrinogen and MTHFR variants. 
 Conclusions: It is clear that the PAI-1 4G allele and the 4G/4G genotype have a significant contribution to the development of coronary artery disease and ultimately myocardial infarction. Prophylactic treatment should be considered in patients with this variant.

Association of Matrix Metalloproteinase-1 Promoter Polymorphisms With Asthma Risk.

Matrix metalloproteinase-1 (MMP-1) expression has been documented as an influential contributor to the intricate milieu of allergic airway inflammation, tissue remodeling, and the exacerbation of asthma's severity. However, the genetic role underlying MMP-1 in the context of asthma has remained enigmatic, with its full implications yet to be unveiled. Considering this, our research was designed to investigate the association of MMP-1 -1607 rs1799750 and the propensity for asthma severity. As a case-control investigation, our study enrolled 198 individuals diagnosed with asthma and age- and sex-matched 453 non-asthmatic controls. The genotypes of MMP-1 rs1799750 were determined utilizing the polymerase chain reaction-restriction fragment length polymorphism methodology. The frequency distributions of 2G/2G, 1G/2G and 1G/1G genotypes at MMP-1 rs1799750 were 49, 42.9, and 8.1%, respectively, among the patients with asthma. This pattern was not different from that of controls (43.7, 46.8, and 9.5%, respectively) (p for trend=0.4486). The allelic frequency pertaining to the variant 1G allele within the asthma group was 29.5%, with a non-significant disparity compared to the 32.9% in the control group (p=0.2596). Noticeably, there was a positive association between MMP-1 rs1799750 2G/1G and 1G/1G genotypes with asthma severity (p=0.0060). Our research indicated that the presence of MMP-1 rs1799750 1G allele might not be the sole arbiter of an individual's susceptibility to asthma, yet its potential to function as a discerning prognostic marker for the severity of asthma emerged as a noteworthy finding deserving attention and further exploration.

Association of polymorphic variants of hemostatic system genes with the course of COVID-19

COVID-19 is characterized by a varied clinical course. The aim of the work was to identify associations of SNPs of hemostatic system genes with COVID-19. DNA was isolated from patients (n=117) and healthy participants (n=104). All infected patients were divided into 3 groups, depending on disease severity assessment, which was appreciated by NEWS2. Another group consisted of participants, who had asymptomatic infection in the past. Determination of SNPs of the genes FGB (-455 G/A), FII (20210 G/A), FV (1691 G/A), FVII (10976 G/A), FXIIIA1 (103 G/T), ITGA2 (807 C/T), ITGB3 (1565 T/C), SERPINE1 (-675 5G/4G) were performed by PCR using the "Genetics of Hemostasis" kit ("DNA-Technology", Russia). In analyzed SNPs, no significant differences were detected between the group of infected patients and healthy participants. But significant association was revealed in gene SERPINE1 (-675 5G/4G), when patient groups, differing in the disease severity, were analyzed relative to the group of participants with asymptomatic infection (p=0.0381; p=0 .0066; p=0.0009). It was found, that as COVID-19 severity scores increased, the proportion of 5G allele of gene SERPINE1 decreased, and the proportion of the 4G allele increased (p=0.005; p=0.009; p=0.0005). Similar processes were observed for genotypes 5G/5G and 4G/4G. The gene SERPINE1 (-675 5G/4G) is associated with the severity of COVID-19. For the first time, it was discovered that 5G/5G genotype of gene SERPINE1 (-675 5G/4G) can be a marker of a milder course of COVID-19, and the 4G/4G genotype as a more severe one.

Evaluation of matrix metalloproteinase-1, -2, -3, -7, and -13 gene polymorphisms in patients with chronic periodontitis and healthy controls.

The current study aimed to investigate the association of matrix metalloproteinase- (MMP-) 1, -2, -3, -7, and -13 gene polymorphisms with chronic periodontitis (CP) in an Iranian population. In this case-control study, 87 subjects with CP and 89 periodontally healthy subjects were allocated to case and control groups, respectively. Subjects' venous blood samples (5cc) were collected, and DNA extraction was performed. A spectrophotometer was utilized to assess the concentration of extracted DNAs. The desired gene polymorphisms were examined using restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR) followed by electrophoresis. Statistical analyses were done using the Pearson Chi-Square test, odds ratio, and t-Test using SPSS Version 28. The MMP-1 (-1607 1G/2G) rs1799750, MMP-3 (-1171 5A/6A) rs3025058, and MMP-7 (-181 A/G) rs11568818 gene polymorphisms significantly differed between case and control groups (PV = 0.019, 0.007, and 0.028, respectively). In contrast, the gene polymorphisms of MMP-2 (-1306 C/T) rs243865 and MMP-13 (-77 A/G) rs2252070 did not make a significant difference. Regarding allele frequencies, the presence of the 2G allele in the MMP-1 (-1607) rs1799750 genotype increased the CP susceptibility significantly, while subjects with the 6A allele in their MMP-3 (-1171) rs3025058 genotype showed significantly lower susceptibility to CP (PV = 0.008 and < 0.001, respectively). In the studied population, gene polymorphisms in the DNA sequences of MMP-1 (-1607 1G/2G) rs1799750, MMP-3 (-1171 5A/6A) rs3025058, and MMP-7 (-181 A/G) rs11568818 may have impacts on CP incidence. Clinicians should be cautious about the association between MMP-1, MMP-3, and MMP-7 gene polymorphisms and the incidence of chronic periodontitis during periodontal treatment planning.

The Effect of Genetic Variation on the Sensitivity to Opioid Analgesics in Patients With Postoperative Pain: An Updated Meta-analysis

BACKGROUND: Responsiveness to opioid analgesics differs among patients with acute postoperative pain. OBJECTIVE: Our study presents the most recent evidence on the effect of genetic variations on postoperative pain, opioid consumption, nausea, and vomiting in patients treated with opioids. STUDY DESIGN: An updated systematic review and meta-analysis on the association between single-nucleotide polymorphisms and opioids administered to patients with acute postoperative pain. METHODS: PubMed, Embase, ISI Web of Science, and the Cochrane Library databases were searched for articles published from February 1, 2014, through December 31, 2021. RESULTS: Added to the previous meta-analysis, 39 studies (a total of 7,455 patients) were included in the final meta-analysis. Highlights of the findings include: 1) human mu-opioid receptor gene 118G allele carriers required more opioids during the first postoperative 24 hours (standard mean difference [SMD] = -0.27; 95% CI,-0.40 to -0.14; P &lt; 0.0001) and 48 hours (SMD = -0.52; 95% CI, -0.83 to -0.20; P = 0.001), and reported higher pain scores during the first 24 hours but not at the 48-hour postoperative period (SMD = -0.09, 95% CI, -0.15 to -0.03; P = 0.002) compared to homozygous 118AA patients. 2) patients with the CYP3A4 *1G allele required fewer opioids during the first 24-hour postoperative period (SMD = 0.59; 95% CI, 0.05 to 1.14; P = 0.03) compared to patients with the homozygous CYP3A4*1/*1 allele. 3) Adenosine triphosphate-binding cassette subfamily B member-1 (ABCB1) 3435T allele carriers required more opioids during the 48-hour postoperative period (SMD = -0.21; 95% CI, -0.38 to -0.04; P = 0.02) compared to homozygous CC carriers. 4) Catechol-O-methyl transferase 158A allele carriers required fewer opioids during the first 24-hour postoperative period (SMD = 0.33; 95% CI, 0.15 to 0.51; P = 0.0004) compared to homozygous GG carriers. No significant differences were observed in patients with CYP2D6*10 and ABCB1 G2677A/T genetic polymorphisms. LIMITATIONS: Several loci were not analyzed in detail due to insufficient clinical data. Furthermore, nongenetic factors that affected analgesic efficacy and the clinical outcome of postoperative pain were not discussed and were not the aim of this meta-analysis. CONCLUSIONS: In combination with previous systematic reviews and meta-analyses, our results indicate that the A118G allele variant of OPRM1 and the *1*1G allele variant of CYP3A4 have a profound influence on individual differences in opioid reactivity in patients with postoperative pain. Our results, together with the identification of additional single nucleotide polymorphisms in future studies, may provide a theoretical basis for precise clinical analgesia. KEY WORDS: Single nucleotide polymorphism, postoperative pain, opioid, meta-analysis

Prevalence of PAI-1 675 4G/5G and ACE I/D polymorphic variants in women with recurrent pregnancy losses

Aim. The present study aimed to assess the distribution of inherited risk factors of thrombophilia among women with RPL. To determine the frequency and distribution of alleles and genotypes of polymorphic loci PAI-1 6754G/5G and ACE I/D among women with RPL. Methods. The study was carried out in a group of 97 women (57 controls and 40 women with RPL). Polymerase chain reaction and restriction fragment length polymorphism (RFLP) were used to identify the polymorphisms. Results. The frequency of 4G allele of PAI-1 675 5G/4G locus was more frequent in the group of women with RPL – 67 %, compared to 54 % in the control group. It was established that the presence of the 4G allele increases the risk of RPL by almost 2 times (Р = 0.01). In the group of women with RPL the frequency of carriers of DD genotype and D allele of ACE gene was higher than in the control group (31 % vs. 19 %), but no significant differences were found. The frequency of women with a combination of 4G4G and DD genotypes of the two studied loci was 16.6 % in the group with RPL versus 2.3 % in the control group. It was shown that the presence of homozygotes for two alleles 4G of the PAI-1 gene and D of the ACE gene leads to an 8-fold increased risk of RPL (OR = 8.6, CI: 0.9841-75.154, P = 0.04). Conclusions. Significance of PAI-1 675 4G/5G and ACE I/D polymorphic variants, especially in combination, in the predisposition to RPL was established.

Association of single nucleotide polymorphisms (4G/5G) of plasminogen activator inhibitor-1 and the risk factors for placenta-related obstetric complications.

Placenta-related obstetric complications (PROCs) such as miscarriage, fetal growth restriction, preeclampsia, and preterm birth are the major causes of maternal and fetal morbidity and mortality. The objective of this study was to search the relevance of plasminogen activator inhibitor-1 (PAI-1) polymorphisms and co-morbidities and the risk factors for PROCs such as miscarriage, fetal growth restriction, preeclampsia, and preterm birth. This retrospective study analyzed the PAI-1 genotype in a cohort of 268 multiparous women with poor obstetric history. Poor obstetric history was defined as the presence of at least one of the PROCs and/or poor gestational outcomes at the previous pregnancy/pregnancies. 5G allele frequency was higher than the 4G allele frequency in the cohort (0.767 vs. 0.233). The frequencies of having at least one risk factor are relatively similar among the different PAI-1 genotypes (P > 0.05). However, the presence of MTHFR polymorphisms (homozygous and compound heterozygous forms of C677T and A1298G) and hereditary thrombophilia (Factor V Leiden and prothrombin G20210A gene mutations, and FXIII deficiency) were found to be associated with PAI 4G/4G (P = 0.048) and 5G/5G (P = 0.022) genotypes, respectively. Significant differences were not observed in other risk factors and co-morbidities such as autoimmune disorders, chronic inflammatory diseases, history of venous thromboembolism, carbohydrate metabolism disorders, hyperlipidemia, cardiovascular and cerebrovascular diseases depending on PAI-1 genotypes (P > 0.05). MTHFR polymorphisms were found to be associated with PAI 4G/4G genotype, while 5G/5G genotype was observed more frequently in hereditary thrombophilia cases.

Association of Plasminogen Activator Inhibitor-1 4G/5G and Angiotensin-Converting Enzyme I/D Polymorphisms with Recurrent Pregnancy Loss in Sudanese Women: A Case-Control study

The aim of our study was to investigate the relationship between the ACE I/D and PAI-1 4G/5G polymorphisms and RPL in Sudanese women. Methods and Results: A total of 232 people participated in this case-control study, including 119 women who had been diagnosed with RPL (Case group) and 113 healthy women (Control group). The case group (RPL) consisted of Sudanese women (mean age of 31.3+5.9 years) who had at least three unfavorable pregnancy outcomes. Women in the control group (Control) were matched by age (mean age of 30.3+5.4 years), had at least two healthy pregnancies, and had no history of unfavorable pregnancy outcomes or recurrent losses. Genomic DNA samples were isolated from the whole blood by using the GF-1 Blood DNA Extraction Kit (Vivantis Technologies Sdn. Bhd., Malaysia) according to the manufacturer`s protocol. The status of the PAI-1 4G/5G and ACE I/D polymorphism was determined by PCR. Analysis of the multiplicative and additive models for the ACE I/D polymorphism showed a significant risk of RPL with the carriage of the D allele (OR=2.07, 95% CI: 1.28-3.35, P=0.003) and the homozygous DD genotype (OR=2.40, 95% CI: 1.34-4.29, P=0.008). The multiplicative and additive models for the PAI-1 4G/5G polymorphism showed a significant risk of RPL with the carriage of the 4G allele (OR=3.11, 95% CI: 2.12-4.58, P=0.000) and the homozygous 4G/4G genotype (OR=3.09, 95% CI: 1.77-5.39, P=0.000). However, the carriage of risk-polymorphic markers, the ACE I/D and PAI-1 4G/5G polymorphisms, was not associated with the number of RPL. The combined carriage of the homozygous DD genotype and heterozygous ID genotype of the ACE I/D polymorphism with the homozygous 4G/4G genotype of the PAI-1 4G/5G polymorphism occurs significantly more often in RPL women than healthy women (P=0.000 and P=0.006, respectively). Carriage of the PAI-1 5G/5G genotype in healthy women was not associated with the ACE I/D polymorphism. Conclusion: Testing for the ACE I/D and PAI-1 4G/5G polymorphisms should be part of the standard examination for patients with RPL.

Analysis and evaluation of correlation between DNA polymorphism in the genes MTHFR, PAI-1 and serum creatinine, creatinine clearance and albumin/creatinine ratio in morning urine of patients with type 2 diabetes mellitus and diabetic nephropathy.

Diabetic nephropathy is a major microangiopathic complication of type 2 diabetes and a leading cause of chronic kidney disease (CKD).

Association of Celiac Disease and Plasminogen Activator Inhibitor-1 Polymorphism

Objective: Celiac disease (CD) is an autoimmune disease. Although susceptibility to thrombosis in celiac patients has been reported in case studies, the cause is not yet known. PAI-1 inhibits fibrinolysis. In this study, the association between celiac disease and PAI-1 4G/5G polymorphism was investigated among celiac patients whose disease were confirmed by intestinal biopsy and healthy controls. Methods: Biopsy-diagnosed celiac patients (n=56) and healthy controls (n=52) were included in the study. PCR-RFLP method was used for PAI-1 genotyping and the results were compared according to statistical significance. Results: The PAI-1 4G allele associated with thrombotic risk and inflammation was found to be higher than the 5G allele in cases (0.63 and 0.36, respectively, p=0.054). When the age at diagnosis was compared according to the PAI-1 variants, no significant difference was found (p=0.3). Although the genotype frequencies were similar in 4G/4G patients and controls, the 5G/5G genotype, known as the protective genotype, was found to be higher in controls (35% and 18%). Conclusion: In our study, the 4G allele of PAI-1, which plays a role in the susceptibility to thrombosis, was observed more frequently in celiac patients compared to the control group. Studying genetic markers of thrombosis in celiac patients is important for individual prophylaxis.

Family-based analysis of -675 4G/5G polymorphism in the PAI-1 gene of polycystic ovary syndrome in Chinese population

Previous studies have shown that 4G/5G polymorphism in promoter region of plasminogen activator inhibitor-1 (PAI-1) gene can affect insulin sensitivity by elevating the level and activity of plasma PAI-1. In order to elucidate the relationship between the polymorphism of PAI-1 gene and polycystic ovary syndrome (PCOS), we used transmission disequilibrium test (TDT) to study the family of PCOS. Eight hundred and fifty-five participants consisting of 285 trios (mother, father and offspring with PCOS) were recruited at the Center of Reproductive Medicine, Shandong University from July 2007 to August 2014. 4G/5G polymorphism of PAI-1 gene was genotyped using direct sequencing protocol and TDT was used to analyse the association between PAI-1 gene and PCOS. Though the 5G allele in PAI-1 gene was overtransmitted in families, no statistical significance existed and there was no association between PAI-1 gene and PCOS, indicating that PAI-1 gene was unlikely to play a major role in the aetiology of PCOS in Chinese population. Impact Statement What is already known on this subject? Some studies have shown that 4G/5G polymorphism in promoter region of plasminogen activator inhibitor-1 (PAI-1) gene can affect insulin sensitivity by elevating the level and activity of plasma PAI-1, participating in the formation of insulin resistance (IR). What do the results of this study add? Though the 5G allele in PAI-1 gene was overtransmitted in families, no statistical significance existed and there was no association between PAI-1 gene and polycystic ovary syndrome (PCOS). What are the implications of these findings for clinical practice and/or further research? PAI-1 gene was unlikely to play a major role in the aetiology of PCOS.

The Role of 4G allele in Plasminogen Activator Inhibitor type-1 rs1799889 Gene as Biomarker to Thrombophilic Complication among Type 2 Diabetic Patients

The Role of 4G allele in Plasminogen Activator Inhibitor type-1 rs1799889 Gene as Biomarker to Thrombophilic Complication among Type 2 Diabetic Patients

Polymorphism of genes of hemostasis system and methionine exchange in patients with female reproductive tumors.

e17500 Background: Our purpose was to analyze the rates of polymorphic allelic variants of genes of hemostasis system and methionine exchange in patients with female reproductive tumors. Methods: The study included 51 patients with histologically verified gynecologic tumors (group 1), including 28 patients (group 1a) with malignant tumors (cervical cancer (CC) n = 8, ovarian cancer (OC) n = 8, endometrial cancer (EC) n = 8, other cancers n = 4) and 23 patients (group 1b) with benign tumors, and 47 women without tumors (group 2). 12 polymorphic loci were studied by RT-PCR in genomic DNA samples: F2 (G20210А, rs1799963), F5 (G1691A, rs6025), F7 (G10976A, rs6046), F13 (G226A, rs5985), FGB G(-455)A (rs1800790), ITGA2-α2 (C807T, rs1126643), ITGB3-b (Т1565С, rs5918), PAI-1 4G(-675)5G, rs1799889), MTHFR (С677Т, rs 1801133 and A1298C, rs1801131), MTR (А2756G, rs1805087), MTRR (A66G, rs1801394). Groups 1, 1a and 1b were compared with controls (p1) and among themselves (p2). Results: The ratio of genotype frequencies maintained in the Hardy-Weinberg equilibrium in all gene loci except F7 (G10976A) in group 1 (p = 0.03). An alternative allele in the F2 gene was found only in group 2 (1.1%). The frequency of an alternative allele in the F5 gene in group 1 was 2.9%, including 1a – 1.8%, 1b – 4.3%, group 2 – 2.1%; F7 – 16.7%, 14.3%, 19.6% and 17.0%; F13 – 23.5%, 23.2%, 23.9% and 34%; FGB – 26.5%, 25.0, 28.3% and 25.5%; ITGA2 – 53.9% (p1= 0.03, OR = 1.89 (1.07-3.33), 48.2%, 60.9% (p1= 0.01, OR = 5.21 (1.22-5.17) and 38.3%; ITGB3 – 13.7%, 10.7%, 17.4% and 16.0%; PAI-1 – 47.1% (p1= 0.03, OR = 0.53 (0.30-0.93), 46.4%, 47.8% and 62.8%; MTHFR (Т) – 28.4%, 30.4%, 26.1%, 34.0%; MTHFR (С) – 34.3%, 28.6%, 41.3% (p1= 0.04, OR = 2.17 (1.02-4.61) and 24.5%; MTR – 18.6%, 19.6%, 17.4% and 27.7%; MTRR – 63.7%, 71.4%, 54.3% and 62.8%, respectively. TT genotype at the ITGA2-α2 (C807T) locus was more frequent in group 1 than in group 2 (23.5% vs 19.1%, p1= 0.01, OR = 6.54 (2.61-16.40); CT genotype was more frequent in group 1a than in group 2 (67.9% vs 38.3%, p1= 0.004, OR = 3.40 (1.27-9.13), and more frequent in EC than in group 2 (87.5% vs 38.3%, p1= 0.03, OR = 11.28 (1.28-99.40). GG genotype at the MTRR (A66G) locus was more frequent in group 1a than in group 1b (53.6% vs 26.4%, p2= 0.042). 5G5G genotype at the PAI-1 4G(-675)5G locus was more frequent in group 1 than in group 2 (31.4% vs 10.6%, p1= 0.04, OR = 3.84 (1.28-11.53), and more frequent in OC than in group 2 (75% vs 11%, p1= 0.0001, OR = 25.50 (3.96-160.20). AA genotype at the F7 (G10976A) locus was more frequent in CC patients than in group 2 (31.3% vs 17%, p1= 0.03, OR = 15.33 (1.20-195.75). Conclusions: Carriage of the AA genotype at the F7 (G10976A) locus may increase the risk of developing CC, and the CT genotype at the ITGA2-α2 (C807T) locus may increase the risk of EC. On the contrary, the alternative 4G allele at the PAI-1 4G(-675)5G locus was less common in patients with malignant tumors, especially OC, than in the group without cancer.

The Role of 4G Allele in Plasminogen Activator Inhibitor-1 (rs1799889) Gene as Biomarker for Thrombophilic States among Patients with Type 2 Diabetes

The Role of 4G Allele in Plasminogen Activator Inhibitor-1 (rs1799889) Gene as Biomarker for Thrombophilic States among Patients with Type 2 Diabetes

Determination of the prevalence of thrombogenic risk factors on the examination of children in the Health Centre

Introduction. Thrombosis in children and adults is believed to be always multifactorial and caused by a combination of permanent and temporary thrombogenic risk factors. Carriage of hereditary or life-long thrombogenic risk factors accompanying a person, causing a tendency to occur arterial and venous thrombosis is a critical problem, but little studied in paediatrics. Materials and methods. A genetic study of twelve prothrombotic polymorphic variants of candidate genes was carried out at the Health Centre. Three hundred 96 children were examined; 177 (43.3%) were boys, and 219 (56.7%) were girls. The average age of the patients was 15.6 ± 1.8 years. The main group of children with thrombogenic risk factors consisted of 27 cases, including ten boys and 17 girls. The comparison group consisted of 396 adolescents, including 167 boys and 202 girls. Results. The study of genetic polymorphisms of the genes of blood clotting factors and genes of folate metabolism in the work of the Health Centre revealed the prevalence of the A2756G allele of the MTR gene (p = 0.032) and the C1565 allele of the ITGB3 gene (p = 0.012) in the population of girls to be statistically significantly higher than in boys. The proportion of the 4G allele (-675) of the PAI-1 gene (p = 0.028) was also determined to be significantly more often detected in boys, while the proportion of the 5G allele of the PAI-1 gene (p = 0.032) was found with a higher frequency in girls. The distribution of allele and genotype frequencies in the studied genes of blood coagulation factors and folate metabolism were checked for compliance with the Hardy-Weinberg equilibrium. The group of children with thrombogenic risk factors included 27 patients. In children of this group, in 64.3% of cases, a mutation of the GA genotype of the factor V Leiden gene was detected, and 37.5% of children were with compounds from the homozygous TT allele of the C677T MTHFR genotype and the homozygous AA allele of the G(-455)A genotype of the fibrinogen FBG gene. Conclusion. With the introduction of personalized prophylaxis in the thrombogenic risk group based on the children’s health centre, thrombogenic complications can be successfully prevented.

Effect of CYP3A5 and CYP3A4 Genetic Variants on Fentanyl Pharmacokinetics in a Pediatric Population.

Fentanyl is an anesthetic/analgesic commonly used in surgical and recovery settings. CYP3A4 and CYP3A5 encode enzymes, which metabolize fentanyl; genetic variants in these genes impact fentanyl pharmacokinetics in adults. Pharmacokinetic (PK) studies are difficult to replicate in children due to the burden of additional blood taken solely for research purposes. The aim of this study is to test the effect of CYP3A5 and CYP3A4 genetic variants on fentanyl PKs in children using opportunistically collected samples. Fentanyl concentrations were measured from remnant blood specimens and dosing data were extracted from electronic health records. Variant data defining CYP3A4*1G and CYP3A5*3 and *6 alleles were available from prior genotyping; alleles with no variant were defined as *1. The study cohort included 434 individuals (median age 9months, 52% male subjects) and 1,937 fentanyl concentrations were available. A two-compartment model was selected as the base model, and the final covariate model included age, weight, and surgical severity score. Clearance was significantly associated with either CYP3A5*3 or CYP3A5*6 alleles, but not the CYP3A4*1G allele. A genotype of CYP3A5*1/*3 or CYP3A5*1/*6 (i.e., intermediate metabolizer status) was associated with a 0.84-fold (95% confidence interval (CI): 0.71-1.00) reduction in clearance vs. CYP3A5*1/*1 (i.e., normal metabolizer status). CYP3A5*3/*3, CYP3A5*3/*6, or CYP3A5*6/*6 (i.e., poor metabolizer status) was associated with a 0.76-fold (95% CI: 0.58-0.99) reduction in clearance. In the final model, expected clearance was 8.9 and 6.8 L/hour for a normal and poor metabolizer, respectively, with median population covariates (9months old, 7.7kg, low surgical severity).