Thrombophilia Markers Research Articles

Heritable Thrombophilia and Increased Risk for Venous Thromboembolism Despite Thromboprophylaxis After Pelvis or Acetabulum Fracture.

Individuals with pelvic and acetabular fractures are at high risk of venous thromboembolism (VTE). The purpose of this study was to determine whether serum markers for thrombophilia and rapid thromboelastography (r-TEG) values correlate with increased VTE risk among patients with pelvic and acetabular fractures. . Prospective observational study. Two urban academic level 1 trauma centers. Adult patients with isolated pelvis and/or acetabulum fractures (OTA/AO 61 and 62) treated surgically placed on a standardized VTE chemoprophylaxis regimen with enoxaparin over a 5-year period were included. Serum r-TEG, coagulation laboratory values, and markers for heritable thrombophilia were drawn postoperatively and after completion of a 6-week course of enoxaparin. The primary outcome was VTE event (either deep venous thrombosis or pulmonary embolism) diagnosed using a Duplex ultrasound, chest computed tomography angiogram, or lung ventilation-perfusion ordered based on clinical suspicion of a VTE event. Laboratory markers and values were then compared between patients who went on to have a VTE event and those who did not and patients with and without markers of thrombophilia. One hundred thirty-three adult patients with isolated operative pelvic and/or acetabular fractures were enrolled in this study. The average age of patients at time of injury was 48.3 years (range 18-91). Sixty-seven percent of patients in the study were (n = 90) males. Sixty-three percent of patients (n = 84) completed both clinical and laboratory follow-up. Forty-one percent of patients (n = 54) had 1 or more markers of heritable thrombophilia. Twelve percent (n = 10) of patients who completed follow-up were diagnosed with VTE. Age, sex, and smoking status were not associated with VTE. Patients who developed VTE had a higher body mass index (P = 0.04). Having more than 1 marker of heritable thrombophilia (P = 0.004) and an r-TEG mean amplitude greater than 72 mm postoperatively was positively associated with VTE (P = 0.02). Among patients treated surgically for isolated pelvic and acetabular fractures who received enoxaparin prophylaxis, the presence of more than 1 marker of heritable thrombophilia or r-TEG mean amplitude value greater than 72 mm postoperatively was associated with an increased risk of VTE. Prognostic Level III. See Instructions for Authors for a complete description of levels of evidence.

Can prothrombotic gene variants and Apoa1 rs5069 polymorphism be the predictors of early myocardial infarctions?

We aimed to determine the genetic risk factors in patients aged 45 years and below with a history of early myocardial infarction (MI), compared to individuals over 60 years of age with no history of MI. In this study, we selected different age groups to more clearly distinguish genetic differences. Accordingly, we compared individuals who had experienced MI at an early age with those who were older and had not experienced any cardiovascular events. The patient group consisted of 99 volunteers under the age of 45 with a history of MI, while the control group included 99 volunteers aged 60 and over without a history of MI. MTHFR (C677T, A1298C), Factor V Leiden (G1691A), Prothrombin (G20210A), PAI (4G/5G), Factor XIII (V34L), APOA1 (rs670, rs1799837, rs5069), and APOB were studied using blood samples taken from the patients. In the logistic regression analysis of thrombophilia markers and gene polymorphisms in the patient and control groups, no statistically significant increase was observed in markers other than APOA1 rs5069 gene polymorphism. APOA1 rs5069 gene polymorphism was found to be higher in the patient group than those without this polymorphism. The frequencies of homozygous MTHFR (C677T, A1298C) and heterozygous Factor XIII V34L were higher in the patient cohort compared to the controls. In our study, we found that prothrombotic gene variants and APOA1 rs5069 polymorphism were statistically significantly associated with coronary artery disease. Thus, prothrombotic gene variants and APOA1 rs5069 polymorphism may serve as predictors of early myocardial infarctions. Individuals with early family histories of coronary artery disease could be screened for these mutations.

Comprehensive evaluation of genetic and acquired thrombophilia markers for an individualized prediction of clinical thrombosis in patients with lymphoma and multiple myeloma

Patients diagnosed with lymphoma or multiple myeloma are at elevated risk of venous thromboembolism (VTE). Optimum risk stratification and effective thromboprophylaxis can only be achieved through the development of a multiple-specific risk score that successfully captures all aspects of the heterogeneous prothrombotic environment existing in these patients. Our aim was to identify risk factors for thrombosis and suggest an improved tool combining clinical data, thrombo-inflammatory biomarkers and genetic (Thrombo inCode® test) variables for predicting thrombotic risk in patients with lymphoma and multiple myeloma. A prospective longitudinal study was conducted on newly-diagnosed lymphoma and multiple myeloma patients who presented at our institution between February 2020 and January 2021. The study included 47 patients with lymphoma and 16 patients with multiple myeloma. We performed a follow-up of 1 year or until September 2021. The incidence of venous thrombosis and associated risk factors were analysed, including the genetic Thrombo inCode® test. Khorana and ThroLy scores for lymphoma patients and IMPEDE VTE score for myeloma patients were calculated. At a median follow-up of 9.1 months, VTE incidence was 9.5% (6/63), with 4 and 2 patients with lymphoma and myeloma who developed the events, respectively. Univariate analysis showed that the incidence of thrombosis was significantly higher in patients with ECOG ≥ 2 and prior immobility. Median factor VIII levels were significantly higher in patients with thrombosis (with increased values in all of them). Moreover, there was a trend in genetic variant rs5985 (factor XIII) as a protective factor, and a trend to higher thrombotic risk in patients with factor V Leiden, rs2232698 variant (serpinA10), low total protein S activity, elevated D-dimer, aggressive lymphoma and treatment with dexamethasone. The results of our study demonstrate promise for the potential use of widely accessible markers to increase precision in risk prediction for VTE in patients with lymphoma and multiple myeloma, particularly ECOG ≥ 2, immobility and higher factor VIII levels, as well as lymphoma aggressiveness, treatment with dexamethasone and the haemostatic biomarkers D-dimer and total protein S activity. Additionally, genetic variants factor V Leiden, serpinA10 rs2232698 and factor XIII-A Val34Leu warrant further investigation for use in the research setting.Graphical

Paternal thrombophilia and recurrent implantation failure: an exploratory case-control study.

Many pieces of literature have reported that inherited and acquired thrombophilia might be a risk factor for recurrent implantation failure (RIF), however, most studies have only focused on RIF patients and not their male partners. We studied the possible association of paternal thrombophilia with RIF risk. Forty-two male partners aged 20-45 suffered from RIF compared with 42 males from couples with at least one successful pregnancy. All participants were investigated for thrombophilia markers. The prevalence of coagulation Factor V activity was significantly higher in the case group (42.9%) than in the control group (16.7%) (p=0.008) (OR=3.75; 95% CI, 1.38, 10.12). The prevalence of protein C and protein S deficiencies in RIF patients were 4.8% and 2.4%, respectively, and 0% in the controls. The prevalence of antithrombin III (ATIII) deficiency was significantly higher in the case group (19%) than in the control group (2.4%) (p=0.01). None of MTHFR C677T and MTHFR A1298C were statistically significant between the two groups. Combined thrombophilia was 45.2% in the men of the RIF group when compared with the control, 14.2% (p=0.001) (OR = 4.95; 95% CI, 1.75-13.86). Paternal thrombophilia may be related to recurrent implantation failure, so evaluation of this factor in RIF patients could be used to identify relevant risk groups and may help in the proper management of these cases to enhance the chance of implantation.

Coagulation disorders and thrombophilia markers in a patient with SARS-CoV-2, type 2 diabetes, hypothyroidism and phlebitis of the left pelvic limb

Coagulation disorders and thrombophilia markers in a patient with SARS-CoV-2, type 2 diabetes, hypothyroidism and phlebitis of the left pelvic limb

Features of disorders and methods of correction of the hemostasis system in women with antenatal fetal death

The improvement of the providing medical services quality to pregnant women remains an urgent issue in modern obstetrics. Women with perinatal losses deserve special attention, especially in the second half of pregnancy. Changes in the hemostasis system are an integral part of the development of pregnancy. The tendency to hypercoagulation has significant pathogenetic significance and can be the cause of a number of complications – miscarriage in the I trimester of pregnancy and preeclampsia, premature birth, antenatal fetal death (AFD) – in the II and III trimesters of pregnancy. Also, pregnancy is a background process for the activation of a number of diseases, in particular hereditary thrombophilia.The objective: to evaluate the changes in the hemostasis system in women with antenatal fetal death and the effectiveness of the use of low molecular weight heparins for the correction of disorders in the hemostasis system in this category of patients during childbirth and in the postpartum period.Materials and methods. 72 women were examined, including 42 pregnant women with AFD (main group) and 30 pregnant women with a physiological course of pregnancy, childbirth and the postpartum period (control group). The state of the hemostasis system was studied using a standard coagulogram. D-dimer was determined by immunoturbometric analysis.To assess the state of the hemostasis system, the following biochemical tests were used: procoagulant link – fibrinogen content and indicators: prothrombin index, activated partial thrombin time (APTT), thrombin time (TP), ancistrone time (AT), soluble fibrinogen-monomer complex (SFMC), factor X (FX); to evaluate the antithrombin system, the content of antithrombin-III (AT-III), protein C were determined; to characterize the state of the fibrinolytic system – the amount of plasminogen, α2-antiplasmin, fibrinogen degradation products (FDC).Thrombophilia markers and antiphospholipid antibodies were also determined. The preference was given to vaginal childbirth. During childbirth, mechanical compression of the lower limbs was applied using special compression stockings (compression level 2). In 12 hours after delivery thromboprophylaxis with low molecular weight heparins was started, the drug of choice being enoxaparin sodium. After 48 hours of the postpartum period, a comparative analysis of coagulogram indicators was performed to further determine the timing of thromboprophylaxis.Results. Analysis of family thrombotic anamnesis revealed risk factors in 12 (28.6%) patients of the main group. In close relatives of the first line, the presence of peripheral vein thrombosis was found – 5 (41.7%) cases, myocardial infarction under the age of 45 – 3 (25.0%) cases, pulmonary embolism – 1 (8.3%) case, transient ischemic attack of the brain – 3 (25.0%).In the control group only 2 (6.6%) patients had a family history of venous thromboembolism in first-line relatives. The analysis of the coagulation system shows significantly higher fibrinogen values (the main group – 5.3±0.2 g/l, the control group – 4.3±0.1 g/l; p<0.05), functional FX activity (the main group – 149.3±3.1%, control group – 107.3±2.7%; p<0.05), SFMC (main group – 15.9±1.2 μg/ml, control group – 7.8±0.9 μg/ml; p<0.05) and D-dimer (main group – 4.4±0.25 μg/ml, control group – 0.7±0.2 μg/ml; p< 0.05) in pregnant women of the main group. An increase in these indicators is a predictor of thrombus formation, activation of blood coagulation by the internal pathway with a decrease in the antithrombin reserve due to FX.During the correlation analysis, a strong direct relationship (r=0.8633) was established between the indicators of SFMC and FX in the blood serum of pregnant women of the main group, the combination of which determines the tendency to clot formation. Determination of markers for the most common types of thrombophilia shows the dominance of hereditary forms (prothrombin, Leiden mutation, MTHFR) in 43% of pregnant women of the main group.The implementation of the proposed treatment approach contributed to a significant decrease in the average concentration of fibrinogen 48 hours after delivery in postpartum women of the main group (main group: before delivery – 5.3±0.2 g/l, after 48 hours after delivery – 3.9±0.3 g/l; control group: 48 hours after delivery – 3.2±0.2 g/l; p<0.05) in combination with a synergistic decrease in the average indicators of SFMC (main group: before delivery – 15.9±1.2 μg/ml, 48 hours after delivery – 6.2±0.2 μg/ml; control group: 48 hours after delivery – 5.4±0.3 μg/ml; p<0.05) and FX (main group: before delivery – 149.3±3.1%, after 48 hours after delivery – 103.1±3.6%; control group: after 48 hours after delivery – 117.1±4.1%; p<0.05).Conclusions. Increased coagulation function was confirmed in pregnant women with antenatal fetal death (AFD). Thanks to the proposed method of correcting disorders in the hemostasis system, a decrease in the frequency of postpartum thromboembolism in women with AFD is achieved, as well as an improvement in the main indicators of the hemostasis system. This prevents the emergence of a chronic form of the syndrome of disseminated intravascular blood coagulation which is developed in the cases of AFD.

Online Clinical Decision Support Software Reduces Time to Diagnosis of Fertility Disorders to 8 Weeks on Average [ID: 1380156

INTRODUCTION: Time to diagnosis of fertility disorders is reported to take up to 11 years and involves five to six physicians on average. Prolonged time to diagnosis is not just an immense burden on the health care system, but also means that in vitro fertilization (IVF) becomes necessary for couples who would have been able to conceive naturally had they been diagnosed sooner, often leaving these couples childless. This observational study evaluated how a clinical decision support software based on American Society for Reproductive Medicine, American College of Obstetricians and Gynecologists, and European Society of Human Reproduction and Embryology guidelines used by patients and obstetrician–gynecologists together can reduce the time to diagnosis of up to 65 conditions affecting female fertility. METHODS: One hundred eighty women aged 20–45 who had been trying to conceive for an average of 11 months were recruited to participate. Women completed a reproductive health assessment with a dynamic online questionnaire. The output of the assessment was a lab requisition with an individualized selection of female and male sex hormones, thyroid hormones, markers for glucose metabolism, vitamin deficiencies, complete blood count, as well as markers for immunology, genetics, and thrombophilia. The clinical decision support software combined the results from the online reproductive health assessment with the lab results to render recommendations for diagnosis, further diagnostics, and treatment. RESULTS: An average of four previously unknown health conditions affecting fertility were diagnosed with an average time of 8 weeks to diagnosis (range, 5–16). Diagnoses included polycystic ovarian syndrome, various forms of amenorrhea, premature ovarian insufficiency, vitamin and iron deficiency, hyperthyroidism and hypothyroidism, insulin sensitivity, and prolactinoma. CONCLUSION: An online clinical decision support tool can significantly reduce time to diagnosis of conditions affecting female fertility. It also reduces the number of patient–physician interactions and the time physicians spend on medical history and necessary patient education.

Role of Genetic Thrombophilia Markers in Thrombosis Events in Elderly Patients with COVID-19

Thrombosis is an extremely dangerous complication in elderly patients with COVID-19. Since the first months of the pandemic, anticoagulants have been mandatory in treatment protocols for patients with COVID-19, unless there are serious contraindications. We set out to discover if genetic thrombophilia factors continue to play a triggering role in the occurrence of thrombosis in patients with COVID-19 with prophylactic or therapeutic anticoagulants. We considered the following genetic markers as risk factors for thrombophilia: G1691A in the FV gene, C677T and A1298C in the MTHFR gene, G20210A and C494T in the FII gene, and (-675) 4G/5G in the PAI-I gene. In a cohort of 176 patients, we did not obtain a reliable result indicating a higher risk of thrombotic complications when taking therapeutic doses of anticoagulants in carriers of genetic markers for thrombophilia except the C494T mutation in the FII gene. However, there was still a pronounced tendency to a higher incidence of thrombosis in patients with markers of hereditary thrombophilia, such as FV G1691A and FII G20210A mutations. The presence of the C494T (Thr165Met) allele in the FII gene in this group of patients showed a statistically significant effect of the mutation on the risk of thrombotic complications despite anticoagulant therapy.

Prothrombin Time and Coagulation Factor IX as Hemostatic Risk Markers for Legg- Calvé-Perthes Disease.

Legg-Calvé-Perthes disease (LCPD) is a pediatric disorder that occurs due to the avascular necrosis of the femoral head and affects the range of motion of the hip in various degrees. Its etiology is still unknown, although it has been associated with coagulation abnormalities, however, the lack of reproducibility in the results in various studies has created a controversy as to whether hemostasis disorders are related to LCPD. On the other hand, there is little information on laboratory studies that could facilitate the diagnosis and treatment of LCPD. Blood and plasma samples were tested from 25 patients with LCPD and 50 healthy controls, matched by sex and age. Cellular markers were evaluated through complete blood count, as well as coagulation times, coagulation factors activity, antithrombotic proteins, and homocysteine concentration. After assessing activity value frequencies in each group, the results showed more significant activity in some of the biological risk markers of thrombophilia, presenting a substantial difference in prothrombin time↘, FV↗, FVIII↗, FIX↗, and Hcy↗. These values imply that there may be hypercoagulable states in patients, which can cause thrombotic events. Diminished prothrombin time and increase in FV activity, FVIII, FIX, and Hcy concentration support the hypothesis that microthrombi formation in small-caliber vessels could be causing avascularity and femoral necrosis, which are traits of LCPD. In addition, based on our results, we believe that the laboratory studies carried out are very useful in the diagnosis and treatment of LCPD.

STANDARDIZATION OF THE PROTEIN CALIBRATORS ISOLATION METHODOLOGY FOR THROMBOPHILIA MARKERS DETECTING IMMUNODIAGNOSTIC TEST SYSTEMS

The most accurate laboratory methods for thrombophilia diagnostics are based on the quantitative determination of the blood plasma specific markers that appear as a result of the coagulation cascade activation. Soluble fibrin and D-dimer belong to the main of the last ones. An alteration in the concentration of such markers can indicate thrombin concentration growth and the formation of soluble oligomeric fibrin. It should be pointed out that simultaneous detection of these markers can establish the correlation between the accumulation of soluble fibrin and fibrinolysis and nowadays is provided only by enzyme-linked immunoassay. Thus, the usage of immunodiagnostic test systems for the detection of thrombophilia markers is highly relevant today. The important components of immunodiagnostic test system are protein calibrators, the isolation standardization of which plays a key role for accurate construction of a calibration curve and obtaining objective results as a consequence. Aim. The objective of this study was to develop the soluble fibrin and D-dimer isolation methodology and its standardization for their further use as the protein calibrators for thrombophilia markers detecting immunodiagnostic test systems. Materials and Methods. Soluble fibrin and D-dimer were isolated from collected human blood by fibrinogen salting out with further fibrin polymerization with thrombin and hydrolysis with plasmin. Quality control of the obtained proteins was carried out using SDS-PAGE and turbidimetric measurements with further checking of the proteins as calibrators for the thrombophilia markers detecting immunoassay. Results. Obtained proteins meet the necessary specifications and can be used as calibrators for immunodiagnostic test systems. Soluble fibrin and D-dimer were checked by SDS-PAGE for the absence of impurities. Turbidimetric measurements showed the polymerization capability of the soluble fibrin and the inhibition of the polymerization by D-dimer. Conclusion. The standardized isolation methodology of soluble fibrin and D-dimer can be used to obtain protein calibrators for appropriate immunodiagnostic test systems.

Markers of Hereditary Thrombophilia with Unclear Significance.

Thrombophilia leads to an increased risk of venous thromboembolism. Widely accepted risk factors for thrombophilia comprise deficiencies of protein C, protein S, and antithrombin, as well as the factor V "Leiden" mutation, the prothrombin G20210A mutation, dysfibrinogenemia, and, albeit less conclusive, increased levels of factor VIII. Besides these established markers of thrombophilia, risk factors of unclear significance have been described in the literature. These inherited risk factors include deficiencies or loss-of-activity of the activity of ADAMTS13, heparin cofactor II, plasminogen, tissue factor pathway inhibitor (TFPI), thrombomodulin, protein Z (PZ), as well as PZ-dependent protease inhibitor. On the other hand, thrombophilia has been linked to the gain-of-activity, or elevated levels, of α2-antiplasmin, angiotensin-converting enzyme, coagulation factors IX (FIX) and XI (FXI), fibrinogen, homocysteine, lipoprotein(a), plasminogen activator inhibitor-1 (PAI-1), and thrombin-activatable fibrinolysis inhibitor (TAFI). With respect to the molecular interactions that may influence the thrombotic risk, more complex mechanisms have been described for endothelial protein C receptor (EPCR) and factor XIII (FXIII) Val34Leu. With focus on the risk for venous thrombosis, the present review aims to give an overview on the current knowledge on the significance of the aforementioned markers for thrombophilia screening. According to the current knowledge, there appears to be weak evidence for a potential impact of EPCR, FIX, FXI, FXIII Val34Leu, fibrinogen, homocysteine, PAI-1, PZ, TAFI, and TFPI on the thrombotic risk.

Targeting Thrombophilia in Sickle Cell Disease with Isoquercetin: Results of a Fixed Dose, Single Center, Phase II Randomized Clinical Trial

Introduction: Sickle cell disease (SCD) is characterized by elevated markers of hemostatic activation and thrombophilia manifesting as arterial stroke and venous thromboembolism (VTE). Anticoagulation for VTE in SCD is associated with an increased risk for bleeding. In cancer patients at high risk for VTE, the flavonoid Isoquercetin (IQ) lowered D-dimer and soluble P-selectin possibly by inhibiting plasma protein disulfide isomerase. In a murine model of SCD, IQ with Hydroxyurea (HU) reduced platelet accumulation and fibrin deposition following laser-induced injury. Given these results, we tested the hypothesis that IQ would be relatively safe and reduce thrombo-inflammatory biomarkers in steady state SCD patients. Methods: We conducted a phase II randomized, double blind, placebo-controlled study at the NIH Clinical Center. Eligible patients had SCD, were 60 days remote from an acute VOC or blood transfusion and were on a stable dose of HU. Study subjects (n=46) were randomly allocated to receive either Isoquercetin 1000 mg daily (n= 23) or placebo (n= 23) for 28 days and were monitored at the beginning and at completion of study. The primary endpoint of the study was the ELISA measured change in plasma sP-selectin levels after 28 days of placebo or study drug exposure. Analysis was performed on an intention to treat principle. D-dimer levels and Thrombin anti-thrombin (TAT) complex were measured by Immunoturbidimetric assays and ELISA. Statistical analyses was performed using analysis of covariance (ANCOVA) and Wilcoxon rank-sum test. Results: The mean (± SD) age of the study population was 40 years (± 11.57), and 56% were female. The majority had HbSS disease and were receiving HU therapy. A subset (28%) reported a prior history of venous thrombosis (VTE) and four of these were on extended duration oral anticoagulation (Table 1). Baseline characteristics between the two study groups were relatively similar. For the primary endpoint, at the end of study the effect of IQ on the change in plasma sP-selectin levels after adjusting for the baseline was not significant (Figure 1) (IQ mean change= 0.10 ± 6.53 vs. placebo mean change= 0.74 ± 4.54; p= 0.66). Since treatment with HU could confound a treatment effect of IQ in a small sample, we compared the change of sP-selectin among those receiving and not receiving HU but did not find any significant difference (p= 0.67). In a sensitivity analysis we did not find an interaction between HU and IQ treatment (p= 0.48). A similar examination for the potential role of VTE history (n= 5 in IQ and n= 8 in placebo arm) as a confounder did not show a VTE effect on change in sP-selectin (p= 0.68) nor an interaction between VTE history and treatment (p= 0.65). In addition to the primary outcome, we explored the effect of IQ treatment on traditional biomarkers of thrombosis. In contrast to studies in patients with cancer, IQ exposure for 28 days did not have a noticeable effect on change from baseline in either D-dimer levels (IQ mean change= 0.15 mcg/ml ± 0.92 vs. placebo mean change= -0.10 mcg/ml ± 2.11; p= 0.82) or thrombin-antithrombin (TAT) complexes (IQ mean change= 0.09 ng/ml ± 1.60 vs. placebo mean change= -0.05 ng/ml ±1.29; p= 0.57). Conclusion: Short term Isoquercetin exposure in steady state SCD did not appreciably lower plasma sP-selectin but was well tolerated, safe and did not have bleeding side effects. Clinical studies of higher dose IQ with greater exposure duration may yield more measurable effects on antithrombotic activity. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Laboratory Findings, Medical Imaging, and Clinical Outcome in Children with Cerebral Sinus Venous Thrombosis.

Cerebral sinus venous thrombosis (CSVT) is a rare disease, especially in children. Therefore, thrombophilia markers, risk factors, treatment strategy, and MRI, as well as clinical outcome need further investigation to support future diagnostic and therapeutic guidelines for children. We retrospectively identified all children with CSVT treated in our center between January 1, 2000, and December 31, 2015. Risk factors and laboratory findings were investigated. Furthermore, outcome and treatment satisfaction were evaluated using magnetic resonance imaging (MRI) analyses and a modified questionnaire. All 43 patients, who agreed to participate, were treated with therapeutic levels of heparin; 86% of children had an increased risk for thromboembolic events upon onset of CSVT (acute disease: 58.1%, perinatal risk factors: 9.3%, medical intervention/immobility: 14%, chronic disease: 16.3%). Thrombophilia markers showed positive results (e.g., reduced values for protein C/S, factor-V-Leiden mutation) in 58% of children at the time of CSVT diagnosis but dropping to 20.9% over the course of the disease. Forty-two of 43 patients received MRI follow-ups and the outcome showed complete recanalization in 69% of the patients and partial recanalization in 31%. At the onset of CSVT, 88% of patients reported restrictions in everyday life due to CSVT; at follow-up this percentage declined to 18%. Satisfaction with the outcome among parents/patients according to the questionnaire was high with 1.7 (German school grades from 1 to 6). All 42 children with MRI follow-up demonstrated complete or partial recanalization under anticoagulation. This positive result underlines the need for future studies on anticoagulation to optimize therapy regimens of pediatric CSVT.

Genetic Markers for Thrombophilia and Cardiovascular Disease Associated with Multiple Sclerosis.

Multiple sclerosis (MS) is an autoimmune inflammatory disease of the central nervous system (CNS) with an unknown etiology, although genetic, epigenetic, and environmental factors are thought to play a role. Recently, coagulation components have been shown to provide immunomodulatory and pro-inflammatory effects in the CNS, leading to neuroinflammation and neurodegeneration. The current study aimed to determine whether patients with MS exhibited an overrepresentation of polymorphisms implicated in the coagulation and whether such polymorphisms are associated with advanced disability and disease progression. The cardiovascular disease (CVD) strip assay was applied to 48 MS patients and 25 controls to analyze 11 genetic polymorphisms associated with thrombosis and CVD. According to our results, FXIIIVal34Leu heterozygosity was less frequent (OR: 0.35 (95% CI: 0.12–0.99); p = 0.04), whereas PAI-1 5G/5G homozygosity was more frequent in MS (OR: 6.33 (95% CI: 1.32–30.24); p = 0.016). In addition, carriers of the HPA-1a/1b were likely to have advanced disability (OR: 1.47 (95% CI: 1.03–2.18); p = 0.03) and disease worsening (OR: 1.42 (95% CI: 1.05–2.01); p = 0.02). The results of a sex-based analysis revealed that male HPA-1a/1b carriers were associated with advanced disability (OR: 3.04 (95% CI: 1.22–19.54); p = 0.01), whereas female carriers had an increased likelihood of disease worsening (OR: 1.56 (95% CI: 1.04–2.61); p = 0.03). Our findings suggest that MS may be linked to thrombophilia-related polymorphisms, which warrants further investigation.

The association of thrombophilia in women with severe obstetric complications.

Thrombophilia, where multiple genetic and acquired risk factors interact synergistically, are associated with thrombosis and pregnancy-related complications. Despite being studied profusely, an inconsistent association exists between thrombophilia and pregnancy complications. Between 2018 and 2020, ninety-three women with pregnancy complications were enrolled in the study. Twenty-five healthy pregnant women without pregnancy complications reported to the same hospital were also recruited as controls. Blood samples were tested for homocysteine, coagulation studies, and molecular diagnosis included FVL, PTH and MTHFR genes amplified using PCR strip assay (Vienna Lab Diagnostics, Austria). Other thrombophilia screening, including testing for AT, PC, and LA, were done by chromogenic assays (Dade Diagnostica, Munich, Germany). Homocysteine level was determined by fluorescence polarization immunoassay technology (Axsym, Abbot company, Germany). Overall, 29.03% of women with pregnancy complications had thrombophilia relative to 16% in the control group. However, the difference between the case and control groups did not reach a significant level (p=0.1175). Additionally, combined thrombophilia was more prevalent among cases (10.75%) than in the control group (4%). However, the difference did not reach statistical significance (p=0.1046). Our study demonstrated that the frequency of thrombophilia among healthy women was 16%, and among women with pregnancy-related complications, 29%. Relative to control, all measured thrombophilia markers were more frequent in women with pregnancy-related complications except for LA. Including all the studies on the Saudi population in a meta-analysis study could reveal more information about thrombophilia and pregnancy-related complications in our population.

Polymorphisms of F2 (G20210A), F5 (G1691A), F 7 (G10976A), F 13(G13T), FGB, ITGA2, ITGB3, PAI-I genes and its association with thrombotic complications in patients suffering from Takayasu aortoarteritis of Urals population.

Cardiovascular complications, especially thrombotic events, are characteristic for Takayasu arteritis (TA). These events significantly deteriorate the patients' quality of life and cause disability and preterm death. Coagulation factor II (F2, G20210A), coagulation factor V (F5, G1691A, Leiden), coagulation factor VII (F7, G10976A), coagulation factor XIII (F13, G13T), fibrinogen (FGB), platelet alpha subunit of transmembrane receptor for collagens and related proteins (ITGA2), platelet glycoprotein (ITGB3), and plasminogen activator inhibitor-1 (PAI-I) gene polymorphisms coexist with TA, and their pathophysiologic interaction needs to be studied. A total of 43 patients with TA were examined for nucleotides polymorphism in F2 (G20210A), F5 (G1691A, Leiden), F7 (G10976A), F13 (G13T), FGB, ITGA2, ITGB3, and PAI-I genes using polymerase chain reaction. Moreover, 130 sex- and age-adjusted healthy controls without a history of any thrombotic complications were enrolled. Among the patients with TA, there were 34 women aged between 17 and 77 (mean 49, median 49; Q1-Q3, 36-61) years and 9 men aged between 20 and 66 (mean 37.8, median 38; Q1-Q3: 31-45) years. Thrombotic complications were recorded in 22 (51%) patients with TA. Comparison of thrombophilia markers genotypes in patients with TA and healthy controls revealed homozygous and heterozygous mutation in ITGA2 (p<0.0001) and PAI-I genes (p=0.026). The frequency of occurrence of hereditary thrombophilia markers in patients with TA was assessed. Detection of the PAI-I gene mutation was significantly more frequent (p=0.032) in patients with TA with a history of thrombotic events than in those with no thrombosis history. Detection of multiple (more than 4 genes) simultaneous mutations of thrombophilia markers was significantly (p=0.0001) more frequent in patients with TA with a history of thrombotic events. Assessment of hereditary thrombophilia genetic markers reveals additional (genetic) risk markers of thrombotic complications in patients with TA and may help in decision making for antiplatelet and/or anticoagulant treatment in patients with TA to reduce the risk of thrombotic complications.

Thrombophilia identified as a risk factor for thrombogenesis in cancer patients

Aim: to assess a rate and range of genetic and acquired thrombophilia in onco-gynecologic patients with ovarian cancer, uterine corpus cancer and cervical cancer.Materials and Мethods. A prospective controlled cohort non-randomized interventional study was conducted: within the years 2014 to 2020, there were examined 546 women with genital malignancies, divided into 2 groups: group I – 155 cancer patients with former thrombosis, group II – 391 women with female genital cancer without former thrombotic complications. Control group consisted of 137 patients with benign female genital tumors. The spectrum of circulating APA was studied: antibodies to â2-glycoprotein I (â2-GPI), annexin V and prothrombin as well as genetic thrombophilia due to mutations genes encoding factor V Leiden, methylenetetrahydrofolate reductase (MTHFR) including polymorphism in genes for prothrombin, platelet glycoproteins and plasminogen activator inhibitor-1 (PAI-1).Results. It was found that frequency of circulating APA as well as incidence rate of genetic thrombophilia between cancer patients from group I vs. group II significantly differed: APA in group I vs. group II was 86 (55.5 %; p &lt; 0.01) vs. 92 (23.5 %) compared to 7 (5.1 %) in control group. Genetic thrombophilia was dominated in group I by mutated MTHFR (92.9 %), polymorphismin PAI-1 (28.4 %) and platelet glycoprotein (44.5 %) that were significantly higher (p &lt; 0.05) compared to group II and control group. Hence, it allows to suggest that such identified thrombophilia markers are largely associated with a risk of developing thrombotic complications.Conclusion. Detected high percentage of patients with circulating APA and genetic thrombophilia among cancer patients with former thromboembolic complications corroborate a role for genetic and acquired thrombophilia in developing pre-thrombotic condition. Detecting a range of circulating APA and genetic thrombophilia allows to identify patients who might be referred to a high risk of thrombogenesis and require to preventive application of anticoagulant therapy.

HEREDITARY THROMBOPHILIA IN RECURRENT IVF FAILURE

Background: The predominant part of IVF cycles results in embryo transfer, but only about one third of all cycles reach clinically achieved pregnancy. This is evidence that most embryos failed in an early stage of pregnancy achievement. Recurrent implantation failure after IVF procedures emphasize the clinical importance of this crucial step in assisted reproductive technology. Repeated unsuccessful IVF attempts force efforts to investigate the firm mechanism of the implantation and to find approach to increase pregnancy outcome success. Plenty of factors have been recognized to affect either success, or failure rate of IVF embryo transfer. Maternal side factors include age, parity, hormonal levels before stimulation, antral follicles count, endometrial thickness and quality of transformed endometrium. Other factors, having functions in coagulation and fibrinolysis cascades, were found to be connected with the transformation processes in the endometrium during the implantation.Aim of work: The aim from this study is to evaluate the relation between hereditary thrombophilia and recurrent IVF failure. This will be done by comparing the prevalence of thrombophilia factors mutation in patients with repeated IVF failure and in normal population.Methods: Thirty patients with recurrent IVF failure underwent genetic testing of thrombophilia markers by PCR: Factor II prothrombin mutation, Factor V leiden mutation (FVL), and Methylene tetrahydrofolate reductase (MTHFR) mutation.Results: 13.3% were tested positive for factor V mutation G1619A, 56.7% were positive for MTHFR c667t mutation. While in healthy normal Mediterranean population the percentage of mutation were 3.7% for FVL mutation and 11.1% for MTHFR mutation.Conclusion: hereditary thrombophilia may be considered as a factor in recurrent IVF failure, proper management is to be considered.

The association between thrombophilic genes alterations and poor ovarian response in infertile women: a retrospective case-control study

This research aimed to retrospectively investigate the possible association between poor ovarian stimulation and selected thrombophilia markers in Iranian women with infertility. For this study 100 Iranian infertile women, with a history of at least three Assisted Reproduction Technology (ART) failures (50 with a poor ovarian response and 50 with a normal response), referred to Royan Institute were selected. Targeted genetic variation evaluation for Factor V G1691A, F II Prothrombin G20210A, MTHFR C677T, MTHFR A1298C was performed by PCR-RFLP followed by Sanger Sequencing. The association between these variants and the ovarian response was examined. The results showed an association between Factor V G1691A mutation and poor ovarian response. The heterozygosity rate of the FVL was significantly different between poor responders compared with the normal response group (p-value ≤ 0.05). In conclusion screening of this polymorphism can be used as a genetic determinant of ovarian response functioning through a vascular mechanism. A larger study with bigger sample size is recommended. Impact statement What is already known on this subject? Thrombophilia is a multi-genetic disease that is associated with changes in homeostatic mechanisms. Some studies have suggested that thrombophilia has no relationship with poor ovarian response and reduced ovarian reserve in general infertile population undergoing ART. What do the results of this study add? Our results showed a significant association between the FVL heterozygote mutation and poor ovarian response. What are the implications of these findings for clinical practice and/or further research? Screening of FVL polymorphism may be suggested as a predictive test for ovarian stimulation response in infertile women undergoing ART. Further prospective studies with bigger sample size evaluating other thrombophilia markers and ovarian response, as well as further in-vitro studies may help clarify the biological mechanisms behind the effect of the FVL polymorphism on ovarian response, oocyte quality and embryo quality.

The role of genetic markers of thrombophilia in the structure of the causes of placenta-associated complications in pregnant women with thrombocytopenia

Hypothesis/Aims of study: Disorders of the hemostatic system continue to occupy one of the leading places in the structure of the causes of infertility and miscarriage. While being associated with reproductive losses, hemostatic disorders are a significant link in the pathogenesis of placenta-associated pregnancy complications. The consumption of platelets and blood clotting factors can be a consequence of inherited thrombophilia and the cause of adverse clinical outcomes of pregnancy. Women with congenital and acquired thrombophilia are at high risk of developing both thrombotic and gestational complications of pregnancy. The aim of this study was to assess the frequency of genetic markers of thrombophilia in pregnant women with thrombocytopenia and to determine the risk of developing obstetric complications depending on thrombophilia gene polymorphisms in the examined women.&#x0D; Study design, materials and methods: This multicenter prospective study involved 299 pregnant women in the third trimester of pregnancy. Two groups of patients were included in the study: the main group (n = 249) consisted of individuals with thrombocytopenia, whereas the control group (n = 50) comprised women with normal platelet counts during physiological pregnancy. All patients underwent a complete clinical, anamnestic and laboratory examination. To identify thrombophilia gene polymorphisms, molecular genetic blood testing was conducted using a biochip developed jointly in the Laboratory of Prenatal Diagnostics of Congenital and Hereditary Diseases, the Research Institute of Obstetrics, Gynecology, and Reproductology named after D.O. Ott, Saint Petersburg, Russia and the V.A. Engelhardt Institute of Molecular Biology, Moscow, Russia.&#x0D; Results: The data obtained from a comparative analysis of genetic markers of thrombophilia indicate that the incidence of mutations in the GPIa platelet receptor gene in pregnant women with thrombocytopenia was significantly higher than that in pregnant women with normal platelet counts and physiological pregnancy (42.5% vs. 14.7%; p = 0.003). The frequency of polymorphisms in genes responsible for disorders in the fibrinolytic system (PAI-1, FGB) was also significantly higher than that in the control group (76.4% vs. 47.0%, p = 0.001; 45.7% vs. 23.5%, p = 0.030, respectively).&#x0D; Conclusion: The revealed high frequency of polymorphisms in the platelet receptor genes and genes responsible for disorders of fibrinolysis system in pregnant women with thrombocytopenia may cause platelet hyperaggregation and hypercoagulation, while being a significant risk factor for placenta-associated complications during pregnancy. The variety of genetic defects that may lead to an unfavorable clinical outcome of pregnancy dictates the need for further study.