G20210A Gene Research Articles

Role of Prothrombin and Methylenetetrahydrofolate Reductase Gene Polymorphisms as well as Thrombophilia Markers, as Risk Factors for Unexplained Recurrent Miscarriage: A Case-Control Study.

Unexplained recurrent miscarriage (RM) is still an unsolved reproductive health problem. Inherited thrombophilias have been one of the causes. Mutation in genes encoding coagulation proteins, including prothrombin (PT G20210A) and methylenetetrahydrofolate reductase (MTHFR) genes, increase tendency for venous thromboembolism. This study aimed to evaluate association between polymorphisms in prothrombine and MTHFR genes with RM. We also evaluated association between protein C (PC), protein S (PS), antithrombin III (ATIII), and homocystiene with RM. We conducted a case-control study on women with history of miscarriages and healthy controls. Genetic analysis was done using (TaqMan) polymerase chain reaction (PCR) technique and the other tests were performed to check general health indications and thrombophilia markers. In this study, 195 RM group (group I) participants and 90 healthy controls (group II), PC, PS, ATIII deficiency and Hyperhomocysteinemia were in 7.2, 65.6, 9.2, 10.8% of group I respectively, but was 1.1, 7.8, 2.2, 2.2% of group II. PT G20210A showed two in group I were A/G, no A/G in group II, and no AA carrier in the either group. G allele was observed in 99.5% of the group I and 100% of the group II, while A allele was detected in 0.5% of group I. MTHFR C677T gene showed C/T mutation in 33.3% of group I and 32.2% of group II, while T/T mutation was detected in 12.8% of group I and 8.9% of the group II. C allele was found in 70.5% of group I and 75% of group II, while T allele was found in 29.5% of group I and 25% of group II (P=0.269). PT G20210A and MTHFR C677T gene mutations are not correlated with RM in the Egyptian population. However, Egyptian women with RM are strongly associated with hyperhomocysteinemia, PC, PS, and ATIII deficiencies (registration number: NCT03209063).

MTHFR and MTRR gene polymorphisms in patients with chronic hepatitis B virus infections in Zigong, Sichuan Province

Background Chronic hepatitis B virus (HBV) infection is a severe disease affecting the physical and economic well-being of patients. The relationship between polymorphisms in the MTHFR gene and disease progression following HBV infection remains a controversial topic. Aim To study MTHFR and MTRR gene polymorphisms in patients with chronic HBV infections in Zigong, Sichuan Province. Subjects and methods One hundred and ninety-one patients with chronic HBV infections were divided into three groups: the chronic hepatitis B (CHB) group (n = 71), the hepatitis B-induced liver cirrhosis (LC) group (n = 56), and the hepatitis B-related primary liver cancer (PLC) group (n = 64). The gene polymorphisms were detected using the PCR-melt curve method and analysed. Results The distributions of MTHFR C677T (CC: 41.2% vs. 41.8%; CT: 50% vs. 45.5%; TT: 8.8% vs. 12.7%; p = 0.714), MTHFR A1298C (AA: 70.6% vs. 72.7%; AC: 26.5% vs. 25.5%; CC: 2.9% vs. 1.8%; p = 1.000), and MTRR A66G (AA: 58.1% vs. 65.5%; AG: 39.0% vs. 29.1%; 2.9% vs. 5.5%; p = 0.353) genetic polymorphisms did not vary between male and female patients from Zigong. In addition, there were no differences in the distributions of MTHFR C677T (CC: 43.4% vs. 38.8%; CT: 49.1% vs. 48.2%; TT: 7.5% vs. 12.9%; p = 0.444), MTHFR A1298C (AA: 76.4% vs. 64.7%; AC: 20.8% vs. 32.9%; CC: 2.8% vs. 2.4%; p = 0.155), and MTRR A66G (AA: 62.3% vs. 57.6%; AG: 34.0% vs. 38.8%; 3.8% vs. 3.5%; p = 0.353) genetic polymorphisms between the patients <60 and >60 years of age. The distributions of MTHFR C677T (CHB vs. LC, p = 0.888; CHB vs. PLC, p = 0.661; PLC vs. LC, p = 0.926), MTHFR A1298C (CHB vs. LC, p = 0.12; CHB vs. PLC, p = 0.263; PLC vs. LC, p = 0.550), and MTRR A66G (CHB vs. LC, p = 0.955; CHB vs. PLC, p = 0.645; PLC vs. LC, p = 0.355) gene polymorphisms were comparable between the CHB, LC, and PLC groups. Conclusion The distributions of MTHFR and MRRR genetic polymorphisms in the population with HBV infections in Zigong, Sichuan Province did not differ in age and sex. The MTHFR and MRRR genetic polymorphisms were comparable between the CHB, LC, and PLC groups.

Analysis of FOXE1 rs3758249, IRF6 rs2235375, MTRR A66G in Non-syndromic Cleft Lip and Palate among Indonesian Deutero-Malay Population

Non-syndromic cleft lip and palate (NS-CLP) is one of the most common orofacial malformations, with an incidence of 1 in 700 live births worldwide. This study aimed to determine the risk factor for NS-CLP among the Indonesian Deutero-Malay population by analyzing the FOXE1 rs3758249, IRF6 rs2235375, and MTRR A66G polymorphisms. It is a case-control study, using 50 samples of NS-CLP patients and 50 samples of control (for FOXE1 rs3758249 and MTRR A66G), 30 samples of NS-CLP patients, and 30 samples of control (for IRF6 rs2235375). After DNA was extracted, polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) were performed by using restriction enzymes of Mscl (FOXE1 rs3758249) and Nde1 (MTRR A66G), and sequencing was performed for IRF6 rs2235375. The study was done in the Molecular Genetic Laboratory, Faculty of Dentistry Universitas Padjadjaran Bandung, from September 2023 to January 2024. The chi-square test was used with the exact Fisher's alternatives. The results showed that in FOXE1 3758249, A allele (mutant) was found more in control (OR=0.744, p&gt;0.05), in MTRR A66G, G allele (mutant) was found more in NS-CLP (OR=1.267, p&gt;0.05) meanwhile in IRF6 rs2235375, G allele (mutant) (OR=1.710, p&gt;0.05) was found more in NS-CLP. This study concluded that FOXE1 rs 3758249, IRF6 rs2235375, and MTRR A66G genes were not the risk factor for NS-CLP in the Indonesian Deutero-Malay population.

Genetic thrombophilia and antiphospholipid antibodies in women with early and late preeclampsia: a retrospective cohort study

Aim: to study a pattern of genetic and acquired thrombophilia in pregnant women with severe early-onset (eoPЕ) and severe late-onset PE (loPЕ) preeclampsia (PE).Materials and Methods. A retrospective cohort study was conducted from January 2022 to May 2024. A total of 109 pregnant women were examined: group 1 – 45 women with eoPЕ (&lt; 34 weeks of pregnancy), group 2 – 24 women with loPЕ (≥ 34 weeks of pregnancy), group 3 (control) – 40 women with physiologically uncomplicated pregnancy. All pregnant women were examined for lupus anticoagulant (LA) and antiphospholipid antibodies (aPL). The screening test for aPL included the quantitation of IgG/IgM antibodies against cardiolipin, phosphatidylserine, phosphatidylinositol, phosphatidic acid, and β2-glycoprotein 1 in serum or plasma using an enzyme immunoassay. Genetic thrombophilia, homocysteine, and ADAMTS-13 metalloproteinase (a disintegrin and metalloproteinase with thrombospondin type 1 motif, member 13) levels were also determined.Results. Pregnant women with severe PE more often had genetic forms of thrombophilia (mutations in factor (F) V Leiden gene, prothrombin G20210A, and Thr165Met) and a deficiency of natural anticoagulants (antithrombin and protein S) compared to pregnant women in control group. Women in eoPE vs. loPE group were more often found to carry genetic polymorphisms in the plasminogen activator inhibitor-1 (PAI-1) and fibrinogen genes. Also, in the group of pregnant women with eoPE, the circulation of aPL, ADAMTS-13 inhibitor, and elevated homocysteine levels were more common. Pregnant women with loPE were older and more often suffered from hypertension, diabetes mellitus, and excess body weight. No significant differences between eoPЕ and loPЕ groups were found while comparing prevalence of autoimmune diseases, thrombosis in familial history, mutations in FV Leiden gene (heterozygous form), FII prothrombin gene G20210A (homozygous form), FII prothrombin gene Thr165Met (heterozygous form), antithrombin III deficiency, protein S deficiency.Conclusion. Precise causes underlying PE remain unknown, and we are still far from understanding all the molecular, immunological, genetic, and environmental mechanisms that lead to the various clinical manifestations of placental syndromes including PE. However, the study results suggest that the presence of thrombophilic disorders, especially in the fibrinolytic system, and aPL circulation contribute to eoPE pathophysiology or progression.

Determination of Prothrombin Gene G20210A Mutation in Deep Venous Thrombosis among Sudanese Patients - Khartoum State

Background: Deep vein throm¬bosis (DVT) is serious disorders contributing to increased morbidity and mortality worldwide. However, there is, little genetic data from Africa including Sudan. So this study was conducted to investigate the relationship between genetic mutations G20210A in the prothrombin coagulation factor and deep venous thrombosis in Sudanese patients. Methods: This is a cross-sectional study design was carried out in Khartoum state in the period from December 2014 to May 2018. One hundred of patient group and fifty healthy one were enrolled in this study. Mutation was detected using conventional PCR depending on restriction enzyme polymorphism technique. Results: The mutant allele (G/G) was found in 4 (4%) of patients nevertheless, there is no mutant allele was present in control group (P.value=0.152). Moreover, the mixed allele (G/A) was not detected in patients and healthy control. Exclusively, the control group showed only the wild type (A/A). Conclusion: This study noted a few mutant alleles in patients group and only the wild type (A/A) in control one. However, carrying out a genome-wide associated study is recommended to determine relationship between prothrombin gene mutation and frequency of deep vein throm¬bosis in Sudanese population.

Association of thrombophilic genes (MTHFR, MTR and MTRR) polymorphisms and homocysteine level in relation to the increased risk of thrombosis among COVID-19 patients

BackgroundPatients with COVID-19 have an increased risk of thrombosis and coagulopathy. Homocysteine, an amino acid essential to coagulation, is thought to be involved in such conditions, as its level is mediated by the presence of some single nucleotide polymorphisms (SNPs) in certain genes including MTHFR, MTR and MTRR. AimThe current study aimed to assess the significant role of MTHFR (C677T and A1298C), MTR A2756G and MTRR A66G SNPs as risk factors for thrombosis among Egyptian COVID-19 patients and their effect on homocysteine level. Subjects and methodsThis case-control study was carried out on 90 Egyptian COVID-19 cases, divided into 45 non-complicated cases and 45 complicated cases with thrombosis, in addition to 80 healthy control individuals. DNA was isolated from blood samples of all the participants. The genotyping was performed using real-time PCR; in addition, serum homocysteine level was estimated. ResultsThe MTHFR (C677T and A1298C), MTR A2756G and MTRR A66G variants revealed a significant higher risk of thrombosis under different genetic models including the homozygous comparison of the co-dominant (20 % vs. 2.3 %, OR = 15.88, p = 0.007), the dominant (62.2 % vs. 33.3 % OR = 3.29, p = 0.006) and the allelic models (41.1 % vs. 17.8 %, OR = 3.2, p < 0.001) for MTHFR C677T and also the dominant (77.8 % vs. 51.1 %, OR = 3.3, p = 0.008) and the allelic models (52.2 % vs. 31.1 %, OR = 2.4, p = 0.004) for MTHFR A1298C. In addition, the heterozygous comparison of the co-dominant (60 vs. 11.1, OR = 12.7, p < 0.001), the dominant (62.2 % vs. 11.1 %, OR = 13.1, p < 0.001) and the allelic models (32.2 % vs. 5.6 %, OR = 8.08, p < 0.001) for MTR A2756G and the heterozygous and homozygous comparisons of the co-dominant model [(53.3 % vs. 31.1 %, OR = 3.4, p = 0.03) and (13.4 % vs. 2.2 %, OR = 12.0, p = 0.049), respectively], the dominant (66.4 % vs. 33.3 %, OR = 4.0, p = 0.001) and the allelic models (40 % vs. 17.8 %, OR = 3, p = 0.001) for MTRR A66G. Moreover, MTHFR (C677T and A1298C), MTR A2756G, and MTRR A66G variants were also associated with high levels of serum homocysteine (p = 0.045, 0.001, 0.005 and 0.039, respectively). The homocysteine level was related to the disease severity by its correlation with higher LDH (rs = 0.49, p < 0.001), CRP (rs = 0.44, p < 0.001), IL-6 (rs = 0.46, p < 0.001) and D-dimer (rs = 0.66, p < 0.001) levels. The multivariate regression analysis showed that homocysteine and the dominant models of MTHFR (C677T and A1298C), MTR A2756G and MTRR A66G were independent predictors of susceptibility to thrombosis among COVID-19 patients [(OR = 1.02, p = 0.007), (OR = 1.15, p = 0.043), (OR = 1.2, p = 0.024), (OR = 1.42, p < 0.001), (OR = 1.19, p = 0.021), respectively]. ConclusionMTHFR (C677T and A1298C), MTR A2756G, and MTRR A66G gene variants were associated with high homocysteine and D-dimer levels that carried a risk for thrombosis among COVID-19 patients and associated with the disease progression.

Interrelationships among MTHFR gene polymorphisms, MTRR gene polymorphisms, and HBV gene BCP 1762/1764 mutations with disease progression in Chronic hepatitis B virus infection patients

Objective Chronic hepatitis B virus (HBV) infection is a major disease that seriously affects the health of patients. In this paper, the relationship among MTHFR gene polymorphism, MTRR gene polymorphism and 1762/1764 mutation in the BCP region of HBV gene with disease progression in chronic HBV patients was studied. Methods A total of 144 chronic HBV infection patients from January 2021 to June 2022 in the Third People’s Hospital of Zigong City, were included as the study subjects. These patients were divided into hepatitis B primary liver cancer patients group (PLC) in 51 cases, Non-primary liver cancer patients group (Non-PLC) in 93 cases, Non-PLC is also divided into chronic hepatitis B virus carriers (CHC) in 49 cases, hepatitis B Live cirrhosis(LC) in 44 cases. MTHFR (C677T), MTRR (A66G) and MTHFR (A1298C) genes polymorphisms were detected by PCR-dissolution curve. The level of HBV-DNA was quantified by real-time PCR, and the 1762/1764 mutation site in the BCP region of the HBV gene were detected by ARMS-PCR. Data were statistically analyzed using the SPSS statistical software. Results The proportion of HBV mutations in BCP region 1762/1764 in PLC group was 82.4%, which was higher than that in LC group (63.6%) and CHC group (51.0%), and the differences were statistically significant (p < 0.05). There were no significant differences in the distribution of MTHFR C677T, MTHFR A1298C and MTRR A66G polymorphisms among CHC, LC and PLC (p > 0.05). The polymorphism distribution of MTHFR C677T, MTRR A66G and MTHFR A1298C genes in patients with chronic hepatitis B virus infection at different stages (CHC, LC and PLC) showed no gender or age differences between and within groups (p > 0.05). Among the patients with MTHFR 677CT + TT, MTRR 66AG + GG and MTHFR 1298AA genotype, the proportion of HBV mutation in BCP region 1762/1764 in PLC group was higher than that in CHC group and LC group, and the differences were statistically significant (p < 0.05). Folate levels in the PLC group were lower than those in the non-PLC group (CHC and LC patients), and the difference was statistically significant compared with the CHC group (p < 0.05). In different MTHFR C677T and MTRR A66G genotypes, the serum GGT activity were statistically significant between mutant PLC and mutant Non-PLC (p < 0.05). Conclusion MTHFR C677T, MTRR A66G and MTHFR A1298C gene polymorphisms distribution have no gender and age differences in chronic hepatitis B virus infection patients. The mutation of HBV gene BCP region 1762/1764 may be associated with the occurrence and development of liver cancer in patients with chronic HBV infection. Single difference of MTHFR C677T, MTHFR A1298C and MTRR A66G gene polymorphisms may have little effect on the disease progression in patients with chronic HBV infection. MTHFR 677CT + TT, MTRR 66AG + GG and MTHFR 1298AA genotype combined with HBV gene BCP region 1762/1764 mutation may be closely related to the occurrence and development of hepatitis B liver cancer.

Inflammatory Biomarkers in Patients With Type 2 Diabetes Mellitus and Heart Failure With Preserved Ejection Faction.

To verify the relationship between gene polymorphisms of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) with inflammation markers and codependent metabolic variables in patients with type 2 diabetes mellitus and chronic heart failure (CHF). This study included 154 patients (mean age, 69.1±3.2 years). The control group consisted of 47 patients with metabolic syndrome (MS) without CHF; the 2nd group included 56 patients with CHF with preserved ejection fraction (CHFpEF); and the 3rd group consisted of 51 patients with CHF with reduced ejection fraction (CHFrEF). The rs1800629 polymorphism of the TNF-α gene (TNF-α: G308A) was studied in real time by the polymerase chain reaction (PCR) method and the rs1800795 polymorphism of the IL-6 gene (IL-6: 174 G&gt;C) was studied by PCR with the electrophoretic detection. The frequencies of polymorphic alleles were compared with the clinical blood test results, plasma concentrations of C-reactive protein (CRP), TNF-α, leptin, and fibrinogen. Differences between the groups were determined using the F test. Relationships between individual studied parameters were identified using the regression analysis. In most patients, the occurrence of gene polymorphisms was eident as increased plasma concentrations of biomarkers. An association was found between the TNF-α gene polymorphism (G308A) and an increase in plasma TNF-α and between the IL-6 gene polymorphism (174 C&gt;G) and an increase in plasma CRP. In the CHFpEF group, the rs1800629 gene polymorphism was observed in 55% of patients, among whom 93% had increased TNF-α. The rs1800795 gene polymorphism was observed in 82% of CHFpEF patients, among whom 21% had increased CRP. In the CHFrEF group, the G308A transition in the TNF-α gene was observed in 53% of patients; an increase in the respective cytokine was noted in 67% of patients; the IL-6 gene polymorphism 174 C&gt;G was found in 78%, however, only 14% of patients with this polymorphism had also increased CRP. In the control group, the TNF-α G308A gene polymorphism was found in 30% of patients, while an increase in free TNF-α was associated with this polymorphism in 50% of patients; the IL-6 174 C&gt;G gene polymorphism was detected in 78%, while no increase in the CRP level was observed in this group. This demonstrates a high probability of the TNF-α G308A gene polymorphism occurrence in patients with CHF. Inflammatory markers are important predictors of CHF. The most significant predictor was the TNF-α G308A gene polymorphism, which was observed in more than 50% of patients, the majority of whom had an increase in plasma TNF-α.

Genetic factors of thrombosis &lt;i&gt;FII G20210A&lt;/i&gt;, &lt;i&gt;FV G1691A&lt;/i&gt; (&lt;i&gt;Arg506Gln&lt;/i&gt;) in patients with thoracoabdominal malignant tumors

Introduction. Malignant tumor is one of the leading factors of venous and arterial thrombosis. But there is no data on the need for a genetic testing protocol of cancer patients for genetic predisposition thrombotic conditions, despite the fact that a number of polymorphisms of hemostasis genes are considered to be unconditionally proven factors of high cumulative thrombogenic risk, and proteins encoded by these genes are direct links in the cascades of pathological hypercoagulation in neoplastic processes.Aim. To identify groups of high genetic risk of thrombotic complications among patients with malignant thoracoabdominal tumors.Materials and methods. The study included 223 patients with malignant tumors of the lung, stomach, esophagus, operated in the Department of Thoracic Oncology of the N.N. Blokhin National Research Center of Oncology in 2018–2019. The study groups consisted of patients with myocardial infarction (n = 62), ischemic stroke (n = 24), venous thrombosis/ venous thromboembolic complications (n = 40), patients without cardiovascular diseases, but with a family history burdened by cardiovascular diseases (n = 33). The control group included 81 patients.Results. Among patients with malignant tumors of thoracoabdominal localization, a statistically significant difference was determined in the frequency of carriage of the heterozygous genotype FV 1691GA (Arg506Gln) in patients who had a myocardial infarction (χ2 = 4.0; p = 0.046), who had venous thrombosis (χ2 = 4.118; p = 0.043), in the group of patients with burdened with a family history (χ2 = 4.997; p = 0.026) in comparison with the control group. Statistically significant difference in the frequency of carriage of the heterozygous variant of the mutation in the FII G20210A gene relative to the control group, it was determined in the group of patients who had an acute cerebrovascular accident (χ2 = 6.881; p = 0.009) and among patients with a burdened history (χ2 = 7.563; p = 0.006).Conclusion. In order to assess the risk of development and prevention of thrombotic complications in the perioperative period in patients with malignant thoracoabdominal tumors, who have suffered myocardial infarction, ischemic stroke, venous thrombosis/venous thromboembolic complications, as well as patients without cardiovascular pathology, but with thrombotic conditions in relatives of the first degree, it is advisable to perform DNA diagnostics at the prehospital stage to identify of gene polymorphisms FII G20210A and FV G1691A (Arg506Gln).

Thrombophilia genetic mutations and their relation to disease severity among patients with COVID-19.

Patients with COVID-19 infection appear to develop virus-induced hypercoagulability resulting in numerous thrombotic events. The aim of the present study was to determine the relationship between the thrombophilia genes mutations (prothrombin G20210A, factor V Leiden, and methyltetrahydrofolate reductase (MTHFR)) and the severity of COVID-19 patients. Prospective cross-sectional study. One hundred and forty patients (80 adults and 60 children) were included in the current study. They were divided into the severe COVID-19 group and the mild COVID-19 group, with each group comprising 40 adults and 30 children. The patients were assessed for FV R506Q, FV R2H1299R, MTHFR A1298C, MTHFR C677T, and prothrombin gene G20210A polymorphisms. CBC, D-dimer, renal and liver function tests, hs-CRP, ferritin, and LDH were also assessed. Thrombotic events were clinically and radiologically documented. Severe COVID-19 cases were significantly more frequent to have a heterozygous mutation for all the studied genes compared to mild COVID-19 cases (p<0.05 for all). Being mutant to gene FV R506Q carried the highest risk of developing a severe disease course (p<0.0001). Patients with abnormally high D-dimer levels were significantly more frequent to be heterozygous for FV R506Q, FV R2H1299R, and prothrombin gene G20210A (p = 0.006, 0.007, and 0.02, respectively). We concluded that there is an evident relationship between severe COVID-19 and inherited thrombophilia. In the current study, FV R506Q gene mutation carried the highest risk of developing a severe COVID-19 disease course.

КЛИНИЧЕСКИЕ ОСОБЕННОСТИ ОСТРОЙ ЦИТОМЕГАЛОВИРУСНОЙ ИНФЕКЦИИ, СОПРОВОЖДАЮЩЕЙСЯ ТРОМБОТИЧЕСКИМИ ОСЛОЖНЕНИЯМИ, У ИММУНОКОМПЕТЕНТНЫХ ПАЦИЕНТОВ

Objective of the study. To identify and analyse clinical and laboratory features in immunocompetent patients with thrombotic complications in acute cytomegalovirus infection. Development of the algorithm of prognosis and early diagnosis of thrombotic complications in order to reduce the risk of severe course of the disease and lethal outcome. Materials and Methods. The study included 100 immunocompetent adult patients undergoing inpatient observation and treatment with laboratory-proven acute cytomegalovirus infection. All patients had a detailed clinical and epidemiological history for the possible presence of modifiable and/or non-modifiable prothrombotic risk factors; a multifaceted laboratory and instrumental examination was performed. Results. Out of one hundred patients with acute cytomegalovirus infection selected in the study, thrombotic diseases were observed in 7% of cases (in 7 patients). The mean age of the patients was 41 (38; 42) years. Patients were hospitalised on average on 12 (10; 14) days from the onset of the disease. The main complaints on admission were: prolonged fever up to 38°C and marked general weakness. The patients had elevated ALT and AST in 88% and 80% of cases, respectively. The presence of reactive lymphocytes was registered in 57% of cases. Enlargement of liver and spleen was observed in 67% and 70% of cases, respectively. Compared to patients with uncomplicated course of acute cytomegalovirus infection, statistically significant laboratory differences were observed in SRB (p&lt;0.001) and D-dimers (p&lt;0.05) in patients who had thrombotic complications. All patients who had genetic material (DNA) to CMV in the blood had positive tests for rr65 antigenemia. The polymorphism of factor V gene G1691A (FVLeiden) was detected in only one patient, who was a carrier of heterozygous GA allele. Conclusion. Acute cytomegalovirus infection in immunocompetent patients includes a symptom complex consisting of a typical clinical picture (prolonged febrile intoxication, hepatosplenomegaly, elevated hepatic transaminases, CRP and D-dimer levels) and obligatory laboratory detection of the virus (positive PCR and/or Ag pp65 tests for CMV). Cytomegalovirus is an independent and reliable prothrombotic risk factor, with thrombotic complications occurring in 7% of cases. Patients with risk factors for thrombotic complications with verified acute cytomegalovirus infection are recommended to be prescribed anticoagulant therapy at prophylactic doses. Also, patients with a proven case of thrombotic process on the background of acute cytomegalovirus infection are recommended a course of etiotropic antiviral therapy.

Level of homocysteine and polymorphism of genes involved in folate metabolism in women with polycystic ovary syndrome

Background. Polycystic ovary syndrome (PCOS) is a multifactorial disease in the development of which gene polymorphism plays an important role. In recent years, data on the role of homocysteine (Hcy) in the formation of PCOS have appeared, and hyperhomocysteinemia is even considered one of the main symptoms of this disease. The causes of an impaired Hcy metabolism are varied and mainly depend on the condition of the genes encoding enzymes of the folate cycle. At the same time, available data on the effect of the 5,10-methylenetetrahydrofolate reductase (MTHFR), methionine synthase (MTR), and methionine synthase reductase (MTRR) gene polymorphisms on the development of hyperhomocysteinemia and the risk of PCOS are few and contradictory. The purpose of the study was to investigate the polymorphisms of the main genes encoding enzymes of the folate cycle (MTHFR, MTR, MTRR) and to reveal their relationship with the level of Hcy in PCOS. Material and methods. One hundred and twenty-nine women aged 20–28 years were examined: the main group — 98 patients with PCOS, the control group — 31 healthy women. The serum content of Hcy was evaluated and a molecular genetic study was conducted to identify the MTHFR, MTR, and MTRR genes. Results. Polymorphic variants of genes involved in folate metabolism were found in both patients with PCOS and in healthy women. However, serum concentration of Hcy was significantly higher in PCOS. Analysis of the MTHFR C677T polymorphism gene showed that the presence of the mutant T allele was associated with an increased Hcy level (12.9 ± 0.2 μmol/l) and the risk of PCOS (odds ratio (OR) = 1.19; 95% confidence interval (CI) 0.52–2.71). In the presence of two T alleles, the level of Hcy (14.6 ± 0.3 µmol/L) and the risk of deve­loping PCOS (OR = 7.69; 95% CI 0.98–59.87) increased even further compared to the functionally “normal” C677C genotype. There was also an association between the MTHFR gene polymorphism at locus 1298 and PCOS whose strength depended on the number of pathological C alleles and was mediated by Hcy content, although this mutation was accompanied by a less significant increase in the level of Hcy than the mutation at locus 677. Compared to carriers of the homozygous A1298A genotype, the risk of developing PCOS was 5.7 times higher in patients with one C allele, and 7.3 times higher in the presence of two C alleles. The MTRR A66A and A66G genotypes were associated with a significant increase in the level of Hcy compared to that of the control group and were associated with an increased risk of PCOS. The mutant homozygous G66G genotype was more common in the control group and had no significant effect on Hcy concentration. It is not proved that the MTR gene is a candidate gene for the development of PCOS, and its polymorphic variants have a negative effect on the level of Hcy. The combination of MTHFR C677T and A1298C, MTHFR C677T and MTR A2756G, MTR A2756G and MTRR A66G gene mutations are associated with a greater increase in Hcy and the risk of developing PCOS compared to any individual monomutation. Conclusions. The MTHFR gene polymorphism and the synergistic effect of the MTHFR, MTR, MTRR gene mutations can be important genetic determinants for homocysteine levels and the risk of PCOS.

Risk Factors, Clinical Manifestations, Anticoagulation Duration, and Outcomes in Carriers of Severe Inherited Thrombophilia

Severe Inherited thrombophilia comprises deficiencies of antithrombin (AT), protein C (PC), and protein S (PS), while gain-of-function (GoF) variants include homozygosity for factor V Leiden (FVL) or prothrombin gene G20210A mutation (PGM), or double-heterozygosity. Persons with AT, Protein C, or S deficiencies have a heightened risk of developing thrombosis from a young age. Several family members are usually affected, and thrombosis may occur in unusual locations.Homozygote or double heterozygous of GoF variants may not have a family history of thrombosis, and the first thrombotic event may present later in life. The optimal duration and type of anticoagulation and their long-term outcomes for persons with severe thrombophilia require further investigation. This study aimed to compare risk factors, clinical manifestations, type and duration of anticoagulation and clinical outcomes between persons with AT, protein C, and S deficiencies and homozygote or double heterozygous persons with GoF variants. Retrospective evaluation of electronic medical records of persons with severe inherited thrombophilia referred to the Center for Blood Disorders at Weill Cornell Medicine-New York Presbyterian Hospital between January 2009 and December 2022. Severe deficiencies of AT, PC and PS were defined as (AT ≤ 60%, PC ≤ 50% and PS ≤ 40%). Results needed to be confimed in second testing in one month interval. Statistical analysis was performed using descriptive statistics, and chi-square test and Fisher's exact test were applied to compare variables between persons with AC deficiencies and GoF variants. 795 persons with inherited thrombophilia were identified, of those 671 were excluded as they had mild thrombophilia. Of the remaining 124 persons, 17 were eliminated due to absence of confirmatory results. A total of 107 persons were analyzed. There were 47 (44%) persons with anticoagulant deficiencies (AT, PC, and PS) and 60 (56%) homozygotes for FVL, PGM or double heterozygote. Mean age (SD) was 48 (13.6) and 49 (12.2) for anticoagulation deficiency and GoF mutation, respectively (Table 1). Overall risk factors for thrombosis were similar in both groups. A positive family history of thrombosis in a first-degree family member was higher in the anticoagulant deficient group (68%) compared to the GoF mutation (42%) (p=0.008). In addition, persons with anticoagulant deficiency had higher rates of thrombosis before 40 years of age compared to the GoF mutation group, 68% and 26%, respectively (p=0.031). The most frequent anticoagulant prescribed was a DOAC in both groups, with 38% and 30% used for the anticoagulation deficient group and GoF mutation group, respectively. The duration of anticoagulation was similar in both groups with most patients remaining on anticoagulation for more than one year (79% anticoagulant deficient group, 65% GoF mutation group) (p=0.3). Patients with AC deficiency had more recurrent thrombotic events while on anticoagulation compared to GoF variants (17% vs 6.7%) (Table 2). Our results indicate that patients with severe inherited thrombophilia either due to anticoagulant deficiencies or GoF mutations are likely to remain on anticoagulation for long durations and may have recurrent thrombosis despite therapeutic anticoagulation.

Venous thromboembolism associated with severe dyspnoea and asthma in 102 792 adults.

The most recent guideline on acute pulmonary embolism (PE) indicates possible long-term sequelae such as dyspnoea and chronic thromboembolic pulmonary hypertension after a PE event. However, effects on lung function or asthma risk have not been evaluated in the general population. We tested whether individuals with a venous thromboembolism (VTE) encompassing PE and deep vein thrombosis (DVT) have reduced lung function, or greater risks of dyspnoea and asthma using data from 102 792 adults from the Copenhagen General Population Study. Diagnoses of PE, DVT and asthma were collected from the national Danish Patient Registry. Factor V Leiden and prothrombin G20210A gene variants were determined using TaqMan assays. Prevalences of PE, DVT and VTE were 2.2%, 3.6% and 5.2%, respectively. Individuals with VTE had forced expiratory volume in 1 s of 92% predicted compared with 96% pred in individuals without VTE (p<0.001). Individuals with VTE versus those without had adjusted OR (95% CI) for light, moderate and severe dyspnoea of 1.4 (1.2-1.6), 1.6 (1.4-1.8) and 1.7 (1.5-1.9), respectively. Individuals with VTE versus those without had an adjusted OR for asthma of 1.6 (95% CI 1.4-1.8). Factor V Leiden and prothrombin G20210A genotype also associated with increased risk of asthma (p for trend=0.002). Population-attributable fractions of severe dyspnoea and asthma due to VTE were 3.5% and 3.0%, respectively, in the population. Individuals with VTE have worse lung function and higher risks of severe dyspnoea and asthma, and may account for 3.5% and 3.0% of people with severe dyspnoea and asthma, respectively, in the general population.

Evaluation of acquired and hereditary risk factors for the development of thromboembolism in patients with systemic lupus erythematosus.

Although the contribution of antiphospholipid antibodies (aPL) to thrombolembolism in systemic lupus erythematosus (SLE) is well known, there is not enough data on the contribution of various hereditary thrombophilic factors. In this study, we aimed to determine acquired and hereditary thrombophilic factors in adult patients with SLE. A total of 93 SLE patients (87 women and 6 men) were included. Data on clinical, demographic and laboratory characteristics, and disease activity scores (SLEDAI) of the patients were evaluated. The patients were analyzed with a screen, including lupus anticoagulant, anticardiolipin antibodies (aCL), antithrombin III, protein C, protein S, and homocysteine levels; factor V Leiden ( FVL ), methylenetetrahydrofolate reductase ( MTHFR ) and prothrombin G20210A gene mutations. A total of 23 thromboembolic events were reported in 17 (18.3%) of the patients. The frequency of pregnancy complications and SLEDAI scores were significantly higher in SLE patients who had a thromboembolism event ( P < 0.05). Thromboembolism was detected in 12 (32.4%) of 37 patients with positive aPL antibody and 5 (8.9%) of 56 patients with negative aPL antibody ( P = 0.006). In addition, thromboembolism developed in 11 (32.3%) of 34 lupus anticoagulant-positive patients and 6 (10.1%) of 59 lupus anticoagulant-negative patients ( P = 0.012). Moreover, protein C levels were significantly lower in patients who developed thromboembolism ( P < 0.05). Patients with and without thromboembolism were similar in terms of genetic thrombophilia factors ( MTHFR A1298C, MTHFR C677T, FVL and Prothrombin G20210A ) ( P > 0.05). In conclusion, in the current study, some acquired (aPL, lupus anticoagulant and cCL IGG) and hereditary (protein C deficiency) thrombophilic factors were shown to be associated with the development of thrombosis in SLE patients. However, the effect of other hereditary factors on the development of thromboembolism could not be demonstrated. According to the data of this study, genetic screening seems inappropriate in terms of the risk of thromboembolism in patients with SLE.

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.

The study of Associations between TNFα GENE G308A polymorphism and clinical-neurological, neuroimaging, hemodynamic characteristics and cognitive dysfunction in patients with post-infectious encephalopathy

Introduction. Infectious diseases can affect brain function and cause the development of encephalopathy, even if the pathogen does not directly affect the central nervous system. Infections caused by viruses, bacteria, or parasites can lead to a secondary inflammatory response in the brain, commonly known as neuroinflammation, through the action of inflammatory mediators that affect the brain endothelium and parenchyma, and the response of brain cells to these mediators. Neurological consequences associated with infectious diseases are poorly understood. Nowadays, there is no established strategy for the treatment or prevention of neurological damage associated with peripheral infections. Aim of study was: to establish probable associations of the G308A polymorphic variant of the TNFα gene with clinical-neurological, neuroimaging, hemodynamic characteristics and cognitive dysfunction in patients with post-infectious encephalopathy. Material and methods. 128 patients with PIE who were undergoing treatment in the neurological departments of the communal non-profit enterprise "Ternopil Regional Clinical Psychoneurological Hospital" during 2021-2022 were examined. 26 patients underwent molecular genetic analysis. The control group consisted of 12 practically healthy persons, representative in terms of age and sex. All patients met the inclusion criteria for the study. Neuroimaging was performed using multispiral computed tomography (CT) or magnetic resonance imaging (MRI). The state of cerebral blood flow was studied using transcranial duplex scanning (TCI) of intracranial vessels and extracranial brachiocephalic vessels on a Philips HDI device. Research in the cognitive sphere was carried out using the Montreal Cognitive Test (The Montreal Cognitive Assessment, MoCA). The molecular genetic study of the G308A polymorphic variant of the TNFα gene was carried out according to standard protocols developed in the molecular genetic laboratory of the state institution "Reference Center for Molecular Diagnostics of the Ministry of Health of Ukraine". The results. Analyzing the dependence of clinical-neurological syndromes, neuroimaging, hemodynamic characteristics, and cognitive dysfunction on the polymorphic variant G308A of the TNFα gene in patients with PIE, probable differences in the distribution of genotype frequencies were established only for clinical-neurological syndromes (cephalic syndrome, p=0.005 and movement disorder syndrome, p =0.038) and neuroimaging changes (gliosis phenomenon, p=0.026). Regarding the frequency distribution of alleles of the G308A polymorphic variant of the TNFα gene in patients with PIE, a probable predominance of carriers of the A allele among persons with cephalic syndrome compared to persons without cephalic syndrome was found (91.67% vs. 8.33%). Conclusions. Thus, the allelic polymorphism of the TNFα gene affects the course of PIE, which determines the expediency of further research.

Post-viral idiopathic purpura fulminans is associated with inherited thrombophilia and anti-cardiolipin antibodies.

Idiopathic purpura fulminans (IPF) is a rare and severe coagulation disorder, associated with transient anti-protein S (anti-PS) antibodies in the context of post-viral infection such as varicella. Anti-protein S antibodies are frequently found in the context of varicella, in contrast with the rarity of IPF. Other factors such as anti-phospholipid antibodies (APL) and inherited thrombophilia may be associated with severe vascular complication. This is an ancillary study of a French multicenter retrospective series and systematic review of literature. We analyzed patients who were tested for inherited thrombophilia, namely antithrombin, protein C, protein S deficiency; prothrombin gene G20210A polymorphism (FII:G20210A),Factor V R506Q polymorphism (FV:R506Q); and/or for APL (lupus anticoagulant (LA), anti-cardiolipin antibodies (ACL), or anti-beta 2-GPI antibodies (Aβ2GP1). Among the 25 patients tested for inherited thrombophilia, 7 (28%) had positive results. Three had FV R506Q, two FII:G20210A, one compound heterozygote FV:R506Q associated to FII:G20210A, and one protein C deficiency. APL testing was performed in 32 patients. It was positive in 19 patients (59%): 17 ACL (53%), 5 LA (16%), 4 Aβ2GP1 (13%). The risk of severe complications was not associated with presence of inherited thrombophilia or APL presence, with RR: 0.8 [95% CI: 0.37-1.71], p = 1 and RR: 0.7 [95% CI: 0.33-1.51], p = 0.39, respectively. We found a high prevalence of inherited thrombophilia or APL in a population of patients with IPF. However, we do not find an association with the occurrence of severe vascular complications or venous thromboembolism.

0118 Association between TNF-α G308A Polymorphism (rs1800629) and Insomnia Vulnerability in Fertile Age Women with Primary Dysmenorrhea

Abstract Introduction Higher tumor necrosis factor-alpha (TNF-α) gene expression has been revealed in the peripheral blood monocytes across the menstrual cycle in patients with primary dysmenorrhea (PDM) than in the unaffected control. Sleep quality is affected by environmental factors, endocrine function abnormalities and genetic factors. The aim of this study was to evaluate the effect of TNF-α G308A polymorphism (rs1800629) on the self-reported sleep, health, menstrual pain severity as well as peri-ovulatory serum TNF-α level in women with and without PDM. Methods Sleep, psychological and physiological characteristics and menstrual pain severity were collected with questionnaires and sleep/menstrual diary within a menstrual cycle from a cohort of 43 PDM women and 42 healthy controls (aged 20-30 years, right-handed, non-smoking and non-shift workers without psychiatric, sleep and neurological comorbidity). Chi-square tests were used to check if the genotype proportions of the investigated gene fitted the Hardy-Weinberg equilibrium (HWE) and to evaluate if the genotypic and allelic distribution is associated with PDM. Chi-square tests or analysis of variance were used to examine the effects of genotype, group and/or genotype x group interaction with the sleep, mind and the body health variables. Results Neither allele frequencies nor genotypes of TNF-α G-308A gene could serve as an independent risk factor of PDM in the Taiwanese population. Circulating serum TNF-α did not correspond with the -308 TNF-α promoter polymorphism, either, which might be due to a multifactorial control process. The TNF-α 308 A allele carriers (28.6%) which was less common than the G allele carriers (71.4%), showed lower Beck depression score, lower bodily pain and better physical health and higher subjective sleep quality. Furthermore, in G allele carriers, the PDM group showed higher insomnia severity and lower sleep duration than that of the control group. Conclusion Present results suggested that TNF-alpha polymorphism conferred no prediction to PDM risk and peri-ovulatory serum TNF-α level. However, our findings were evident that the A allele at the TNFα 308 locus, as compared to the more common G allele, was associated to better health and better sleep, with an indication of a genetic underpinning to their better resistance from the risk of PDM. Support (if any)

A Comprehensive Review of Risk Factors for Venous Thromboembolism: From Epidemiology to Pathophysiology

Venous thromboembolism (VTE) is the third most common cause of death worldwide. The incidence of VTE varies according to different countries, ranging from 1-2 per 1000 person-years in Western Countries, while it is lower in Eastern Countries (<1 per 1000 person-years). Many risk factors have been identified in patients developing VTE, but the relative contribution of each risk factor to thrombotic risk, as well as pathogenetic mechanisms, have not been fully described. Herewith, we provide a comprehensive review of the most common risk factors for VTE, including male sex, diabetes, obesity, smoking, Factor V Leiden, Prothrombin G20210A Gene Mutation, Plasminogen Activator Inhibitor-1, oral contraceptives and hormonal replacement, long-haul flight, residual venous thrombosis, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, trauma and fractures, pregnancy, immobilization, antiphospholipid syndrome, surgery and cancer. Regarding the latter, the incidence of VTE seems highest in pancreatic, liver and non-small cells lung cancer (>70 per 1000 person-years) and lowest in breast, melanoma and prostate cancer (<20 per 1000 person-years). In this comprehensive review, we summarized the prevalence of different risk factors for VTE and the potential molecular mechanisms/pathogenetic mediators leading to VTE.