Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 2.265
Filtrar
1.
Vasc Health Risk Manag ; 16: 53-56, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32021228

RESUMO

Introduction: Factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations were investigated in many studies for their association with Deep Venous Thrombosis. Case Presentation: A North Lebanese family has been examined, from an index case, a 40-year-old woman, who had a history of venous thrombosis with unexplained recurrent miscarriage. The index case was found to be heterozygous for factor V Leiden G1691A, prothrombin G20210A, and methylenetetrahydrofolate reductase C677T gene variants. Her family members were heterozygous for at least two of the three-point mutations, and multiple risk factors associated with thrombophilia were identified. Conclusion: Our findings emphasize the need for clarifying the utility and futility of thrombophilia testing in the era of molecular diagnostics.


Assuntos
Aborto Habitual/etiologia , Resistência à Proteína C Ativada/genética , Coagulação Sanguínea/genética , Fator V/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Mutação , Protrombina/genética , Trombofilia/genética , Trombose Venosa/etiologia , Aborto Habitual/sangue , Aborto Habitual/diagnóstico , Resistência à Proteína C Ativada/sangue , Resistência à Proteína C Ativada/complicações , Resistência à Proteína C Ativada/diagnóstico , Adulto , Feminino , Predisposição Genética para Doença , Hereditariedade , Humanos , Líbano , Linhagem , Fenótipo , Gravidez , Fatores de Risco , Trombofilia/sangue , Trombofilia/complicações , Trombofilia/diagnóstico , Trombose Venosa/sangue , Trombose Venosa/diagnóstico
2.
Anticancer Res ; 39(11): 6067-6071, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31704834

RESUMO

BACKGROUND/AIM: Thrombin plays significant roles in various types of cancer. However, the expression levels of prothrombin, the thrombin precursor, in cancer remain unclear. Variants of the 3'end of the prothrombin gene lead to increased prothrombin expression. This study aimed to analyze prothrombin 3'end gene variants in colon tumor and adjacent normal tissue samples. MATERIALS AND METHODS: The study group consisted of 93 patients suffering from colon adenocarcinoma. The 3'end of the prothrombin gene was analyzed by DNA sequencing. RESULTS: Three variants, all previously associated with increased prothrombin expression were detected. Frequency of the FII 19911G allele was 46.77% and 47.85% in tumor and normal tissue, respectively. For the FII 20210A allele, the detected frequencies were 2.15% and 1.61%, respectively. The frequency of the FII c.1824T allele was 0.54% in both tissues. Four patients showed different genotypes in tumor and normal tissue. CONCLUSION: Prothrombin 3' end gene variants may play a role in colorectal cancer.


Assuntos
Regiões 3' não Traduzidas/genética , Adenocarcinoma/genética , Biomarcadores Tumorais/genética , Neoplasias do Colo/genética , Regulação Neoplásica da Expressão Gênica , Variação Genética , Protrombina/genética , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Neoplasias do Colo/patologia , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico
3.
Thromb Haemost ; 119(9): 1441-1450, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31352677

RESUMO

BACKGROUND: The heritability of venous thromboembolism (VTE) is only partially explained by variants in 17 previously VTE-associated genes. OBJECTIVE: This article screens for additional rare variants in the 17 genes and investigates the relative contributions of pro- and anticoagulant genes to VTE. PATIENTS AND METHODS: Ninety-six VTE patients from the population-based Malmö Thrombophilia Study were analysed using an AmpliSeq strategy and Ion Torrent sequencing and the variant data were compared with data from public databases. RESULTS: A total of 102 non-synonymous and 76 synonymous variants were identified. Forty-six non-synonymous variants were present in the human gene mutation database. Anticoagulant and procoagulant genes showed 14 and 22 rare non-synonymous variants, respectively. Individual patients showed varying numbers of risk factors; 13 patients had non-synonymous mutations in SERPINC1, PROC and PROS1 genes and 42 had factor V Leiden or prothrombin mutations generating a total of 47 patients with at least one of these risk factors. Ten common VTE-associated variants showed low level enrichments and no correlation to the other risk factors. The enrichment of previously identified risk factors was similar to previous studies. Determination of the nsyn/syn ratio (number of non-synonymous variants per non-synonymous site, nsyn, to the number of synonymous variants per synonymous site, syn) showed, as expected in patients, an increase of non-synonymous relative to synonymous anticoagulant variants compared with controls (nsyn/syn, 0.95 vs. 0.68). In contrast, non-synonymous procoagulant variants (nsyn/syn, 0.31 vs. 0.63) showed a decrease. We suggest that the deficit of non-synonymous variants in procoagulant genes is a novel mechanism contributing to VTE.


Assuntos
Coagulação Sanguínea/genética , Mutação/genética , Tromboembolia Venosa/genética , Adulto , Idoso , Antitrombina III/genética , Proteínas de Ligação ao Cálcio/genética , Estudos de Coortes , Fator V/genética , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo Genético , Proteína C/genética , Protrombina/genética , Adulto Jovem
4.
Blood Coagul Fibrinolysis ; 30(6): 253-262, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31259774

RESUMO

: A numerous factor can cause infertility, but around one of four reproductive failure cases remain unexplained and diagnosed as idiopathic infertility. In the past few decades, analysis of gene polymorphisms takes a significant place in pathogenesis of infertility. The aim of this study was to evaluate the possible role of hemostasis-related gene polymorphisms in unexplained infertility. The study includes 117 female patients with idiopathic infertility and 130 fertile women with at least one born child. Eight polymorphisms important for hemostasis (ITGB3 1565T>C, FV 1691G>A, FII 20210G>A, MTHFR 677C>T and 1298A>C, ATIII 786G>A, PAI-14G/5G and ACE I/D) were genotyped by real-time PCR system. The frequencies of alleles and genotypes of examined polymorphisms were analyzed in SPSS statistical program, whereas gene interactions were identified using the GMDR software. Examination of etiological factors has shown that family history is a significant factor in assessing individual risk for infertility. The alleles and genotypes frequency of FV 1691G>A and FII 20210G>A polymorphisms were statistically different between control and patient group leading to a greater risk for infertility. The analysis of epistatic relationship between examined hemostasis-related gene polymorphisms identified more complex high-risk genotypes associated with infertility. Our results suggest that positive family history could be important predictive factor for fertility problems, pointing to the potential hereditary basis of this condition. Polymorphisms FVL and FII prothrombin are independent risk factors for idiopathic infertility, whereas multilocus interactions approach should be taken into consideration for the future research.


Assuntos
Epistasia Genética/fisiologia , Hemostasia/genética , Infertilidade Feminina/etiologia , Polimorfismo Genético , Adulto , Alelos , Estudos de Casos e Controles , Fator V/genética , Feminino , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Anamnese , Protrombina/genética , Fatores de Risco
5.
Croat Med J ; 60(3): 212-220, 2019 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-31187948

RESUMO

AIM: To investigate the prevalence of common genetic variants that can serve as markers of thrombophilia and warfarin pharmacogenetics in Bosnia and Herzegovina. METHODS: The study was performed between August and October 2017 on 130 healthy unrelated adult volunteers from Bosnian-Herzegovinian population sample. The prevalence of the following genetic variants was determined: F5 c.1601G>A (factor V Leiden), F2 c.*97G>A (factor II or prothrombin mutation), F13A1 (factor XIII) c.103G>T, MTHFR (methylenetetrahydrofolate reductase) c.665C>T and c.1286A>C, as well as PAI-1 (plasminogen activator inhibitor 1) c.-816A>G and c.-844G>A as markers of thrombophilia risk, and *2 and *3 alleles of CYP2C9 (cytochrome P450 2C9) and five variants of VKORC1 (vitamin K epoxide reductase complex subunit 1) as markers of warfarin pharmacogenetics. DNA was isolated from buccal swabs using salting out method, while genotyping was performed using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. RESULTS: Minor allele frequencies for two main thrombophilia risk factors, F5 c.1601G>A and F2 c.*97G>A were 0.023 and 0.008, respectively. Combined data for the markers of warfarin pharmacogenetics imply that 57.4% study participants can be expected to metabolize warfarin at an extensive, 40.3% at intermediate, and 2.3% at a poor rate. CONCLUSION: This study reports the first extensive population genetic data for thrombophilia and warfarin pharmacogenetic markers in Bosnia and Herzegovina. Allele frequencies of genetic variants are within the general average for European populations, and their presence implies the necessity of introduction of personalized medicine in warfarin-mediated antithrombotic therapy.


Assuntos
Anticoagulantes/metabolismo , Fatores de Coagulação Sanguínea/genética , Trombofilia/genética , Varfarina/metabolismo , Adolescente , Adulto , Alelos , Biomarcadores , Bósnia e Herzegóvina , Citocromo P-450 CYP2C9/genética , Fator V/genética , Fator XIII/genética , Feminino , Frequência do Gene , Genótipo , Humanos , Masculino , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Pessoa de Meia-Idade , Inibidor 1 de Ativador de Plasminogênio/genética , Gravidez , Protrombina/genética , Vitamina K Epóxido Redutases/genética , Adulto Jovem
6.
Braz J Med Biol Res ; 52(4): e8217, 2019 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-30970085

RESUMO

The aim of this study was to perform an updated meta-analysis to quantitatively investigate the association between G20210A polymorphism of Prothrombin gene and the risk of retinal vein occlusion (RVO), based on the available publications with inconsistent results. We utilized the Stata software to perform the heterogeneity test, association test, Begg's and Egger's tests, and sensitivity analysis. We searched three on-line databases (PubMed, Embase, and WOS) and obtained a total of 422 articles. Based on our selection criteria, 24 case-control studies were finally enrolled in this overall meta-analysis; a subgroup analysis by the factors ethnicity, control source, and RVO type was done. Through the association test of overall meta-analysis, we did not observe a significant difference between RVO cases and controls under the A vs G (allele) (z=1.49, P=0.137), A vs G (carrier) (z=1.42, P =0.155), GA vs GG (z=1.50, P=0.135), and GA+AA vs GG (z=1.50, P=0.135). Furthermore, we observed similar negative results in the association test of subgroup analysis (all P>0.05). Heterogeneity, Begg's, and Egger's tests excluded the presence of high heterogeneity and publication bias. Statistically stable results were observed in the sensitivity analyses. Based on integrated analysis of the current evidence, Prothrombin gene G20210A polymorphism is likely unrelated to the risk of RVO.


Assuntos
Predisposição Genética para Doença/genética , Polimorfismo Genético/genética , Protrombina/genética , Oclusão da Veia Retiniana/genética , Genótipo , Humanos , Fatores de Risco
8.
Clin Appl Thromb Hemost ; 25: 1076029619834352, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31025572

RESUMO

Little is known about the pathogenesis of cerebral sinovenous thrombosis (CSVT) in the neonate. Although thrombophilia has been described as increasing the risk of CSVT in adults, it remains controversial in pediatric patients, and prospective case-control studies regarding neonatal CSVT are lacking. From 2008 to 2017, all 26 consecutive newborn infants ≥35 weeks of gestation diagnosed with neonatal CSVT, and their mothers, were tested for factor V Leiden (FV) G1691A, FII G20210A, and methylenetetrahydrofolate reductase C677T (MTHFR C677T) mutations. Eighty-five mother-infant pairs were recruited as controls. All infants except 1 with CSVT were suspected due to clinical symptoms, mainly seizures (22/25). Magnetic resonance imaging was performed in 24/26 infants. Heterozygous FV G1691A, FII G20210A, and homozygous MTHFR C677T mutations were present in 1/26, 3/26, and 3/20 infants with CSVT, respectively. FII (odds ratio: 10.96; 95% confidence interval [CI]: 1.09-110.35) and male sex (3.93; 95% CI: 1.43-10.76) were associated with CSVT. When FII G20210A analysis was adjusted for sex, the OR for FII G20210A was 6.70 (95% CI: 0.65-69.22). No differences were found for FV G1691A or homozygous MTHFR mutations between neonates with CSVT and their mothers, compared to controls.


Assuntos
Fator V/genética , Doenças Genéticas Inatas/genética , Trombose Intracraniana/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Mutação de Sentido Incorreto , Protrombina/genética , Adulto , Feminino , Heterozigoto , Humanos , Recém-Nascido , Masculino , Estudos Prospectivos
9.
PLoS Genet ; 15(3): e1007948, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30870413

RESUMO

Glial cells regulate multiple aspects of synaptogenesis. In the absence of Schwann cells, a peripheral glial cell, motor neurons initially innervate muscle but then degenerate. Here, using a genetic approach, we show that neural activity-regulated negative factors produced by muscle drive neurodegeneration in Schwann cell-deficient mice. We find that thrombin, the hepatic serine protease central to the hemostatic coagulation cascade, is one such negative factor. Trancriptomic analysis shows that expression of the antithrombins serpin C1 and D1 is significantly reduced in Schwann cell-deficient mice. In the absence of peripheral neuromuscular activity, neurodegeneration is completely blocked, and expression of prothrombin in muscle is markedly reduced. In the absence of muscle-derived prothrombin, neurodegeneration is also markedly reduced. Together, these results suggest that Schwann cells regulate NMJs by opposing the effects of activity-regulated, muscle-derived negative factors and provide the first genetic evidence that thrombin plays a central role outside of the coagulation system.


Assuntos
Antitrombina III/genética , Cofator II da Heparina/genética , Junção Neuromuscular/genética , Protrombina/genética , Sinapses/genética , Animais , Perfilação da Expressão Gênica , Camundongos , Neurônios Motores/metabolismo , Neurônios Motores/patologia , Músculo Esquelético/metabolismo , Degeneração Neural/genética , Neuroglia , Junção Neuromuscular/crescimento & desenvolvimento , Células de Schwann/metabolismo , Trombina/genética
10.
J Biol Chem ; 294(19): 7644-7657, 2019 05 10.
Artigo em Inglês | MEDLINE | ID: mdl-30918026

RESUMO

Current thought holds that factor Xa (FXa) bound in the prothrombinase complex is resistant to regulation by protein protease inhibitors during prothrombin activation. Here we provide evidence that, contrary to this view, the FXa-specific serpin inhibitor, protein Z-dependent protease inhibitor (ZPI), complexed with its cofactor, protein Z (PZ), functions as a physiologically significant inhibitor of prothrombinase-bound FXa during prothrombin activation. Kinetics studies showed that the rapid rate of inhibition of FXa by the ZPI-PZ complex on procoagulant membrane vesicles (ka (app) ∼107 m-1 s-1) was decreased ∼10-fold when FXa was bound to FVa in prothrombinase and a further ∼3-4-fold when plasma levels of S195A prothrombin were present (ka (app) 2 × 105 m-1 s-1). Nevertheless, the ZPI-PZ complex produced a major inhibition of thrombin generation during prothrombinase-catalyzed activation of prothrombin under physiologically relevant conditions. The importance of ZPI-PZ complex anticoagulant regulation of FXa both before and after incorporation into prothrombinase was supported by thrombin generation assays in plasma. These showed enhanced thrombin generation when the inhibitor was neutralized with a PZ-specific antibody and decreased thrombin generation when exogenous ZPI-PZ complex was added whether prothrombin was activated directly by FXa or through extrinsic or intrinsic pathway activators. Moreover, the PZ antibody enhanced thrombin generation both in the absence and presence of activated protein C (APC) anticoagulant activity. Taken together, these results suggest an important anticoagulant role for the ZPI-PZ complex in regulating both free FXa generated in the initiation phase of coagulation as well as prothrombinase-bound FXa in the propagation phase that complement prothrombinase regulation by APC.


Assuntos
Coagulação Sanguínea , Fator V/química , Fator Xa/química , Protrombina/química , Serpinas/química , Trombina/química , Substituição de Aminoácidos , Anticorpos/química , Fator V/genética , Fator V/metabolismo , Fator Xa/genética , Fator Xa/metabolismo , Humanos , Cinética , Mutação de Sentido Incorreto , Proteína C/química , Proteína C/metabolismo , Protrombina/genética , Protrombina/metabolismo , Serpinas/genética , Serpinas/metabolismo , Trombina/genética , Trombina/metabolismo
11.
Medicina (Kaunas) ; 55(2)2019 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-30781868

RESUMO

Background and Objective: Evidence indicates that genetic factors may be involved in the risk of ischemic stroke (IS). The aim of this study was to assess the effect of genetic polymorphisms located in exons or untranslated regions of MTHFR as well as FV genes on ischemic stroke. Materials and Methods: In this case-control study, 106 patients with IS and 157 healthy volunteers (age <50 years) were genotyped for MTHFR C677T, A1298C, C2572A and C4869G, FVL, and prothrombin G20210A polymorphisms. Results: The MTHFR 677CT genotype was more frequent in patients and increased risk of IS with Odds Ratio = 1.9. The MTHFR A1298C and C2572A polymorphisms were not associated with IS in dominant and recessive models. Our findings showed a significant decrease in the MTHFR 4869CG genotype in IS patients, and this variant was associated with a decreased risk of IS in the dominant model. The CAAT haplotype was associated with increased risk, and the GAAC haplotype was associated with decreased risk of IS compared to other haplotypes. There was no relation between FVL G1691A polymorphism and IS risk. Conclusions: The present study showed that the MTHFR 677CT genotype was more frequent and the MTHFR 4869CG genotype was less frequent in young IS patients.


Assuntos
Isquemia Encefálica/epidemiologia , Isquemia Encefálica/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Polimorfismo Genético , Adulto , Estudos de Casos e Controles , Fator V/genética , Feminino , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Haplótipos/genética , Humanos , Irã (Geográfico)/epidemiologia , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Protrombina/genética , Risco , Estatísticas não Paramétricas , Inquéritos e Questionários
12.
Biomed Res Int ; 2019: 5653424, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30792993

RESUMO

Most multidrug-resistant tuberculosis (MDR-TB) patients fail to receive a timely diagnosis and treatment. Therefore, we explored the differentially expressed peptides in MDR-TB compared with drug-susceptible tuberculosis (DS-TB) patients using LC-MS/MS and Ingenuity Pathway Analysis (IPA) to analyse the potential significance of these differentially expressed peptides. A total of 301 peptides were differentially expressed between MDR-TB and DS-TB groups. Of these, 24 and 16 peptides exhibited presented high (fold change ≥ 2.0, P < 0.05) and low (fold change ≤ -2.0, P < 0.05) levels in MDR-TB. Significant canonical pathways included the prothrombin activation system, coagulation system, and complement system. In the network of differentially expressed precursor proteins, lipopolysaccharide (LPS) regulates many precursor proteins, including four proteins correlated with organism survival. These four important differentially expressed proteins are prothrombin (F2), complement receptor type 2 (CR2), collagen alpha-2(V) chain (COL5A2), and inter-alpha-trypsin inhibitor heavy chain H4 (ITIH4). After addition of CR2 peptide, IL-6 mRNA expression in THP-1 cells decreased significantly in dose- and time-dependent manners. Cumulatively, our study proposes potential biomarkers for MDR-TB diagnosis and enables a better understanding of the pathogenesis of MDR-TB. The functions of differentially expressed peptides, especially CR2, in MDR-TB require further investigation.


Assuntos
Biomarcadores , Peptídeos/genética , Proteômica , Tuberculose Resistente a Múltiplos Medicamentos/genética , Adulto , Proteínas Sanguíneas/genética , Cromatografia Líquida , Colágeno Tipo V/genética , Feminino , Regulação da Expressão Gênica/genética , Glicoproteínas/genética , Humanos , Masculino , Mycobacterium tuberculosis/patogenicidade , Proteínas Secretadas Inibidoras de Proteinases/genética , Protrombina/genética , Receptores de Complemento 3d/genética , Transdução de Sinais , Espectrometria de Massas em Tandem , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia , Tuberculose Resistente a Múltiplos Medicamentos/patologia
13.
Hamostaseologie ; 39(1): 49-61, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30703819

RESUMO

A thrombophilic disorder is a hereditary or acquired condition that increases the risk of thrombosis. The most common hereditary thrombophilias that predispose to venous thrombosis in the Caucasian population are the heterozygous forms of the factor V Leiden and prothrombin G20210A mutation that are generally detected by direct DNA genotyping. Immunologic antigen assays and chromogenic or clot-based activity assays are used to identify deficiencies in the natural coagulation inhibitors antithrombin, protein C and protein S. Because pre-analytical errors and acquired causes of low antithrombin, protein C or protein S levels are considerably more common than hereditary deficiencies, all potential conditions that may lower activity levels of the natural coagulation inhibitors (e.g. concomitant liver disease, pregnancy, anticoagulant therapy) must be considered and excluded before the diagnosis of an inhibitor deficiency can be made. To avoid misclassification, the diagnosis should not be made based on a single abnormal test result. Thus, repetitive testing when the patient is not on anticoagulant therapy is mandatory to confirm the diagnosis. Screening for antiphospholipid syndrome (APS) comprises testing for lupus anticoagulants (LAs) and the presence of IgG or IgM antibodies directed against phospholipids and phospholipid-binding proteins such as ß-2-glycoprotein-I. A combination of clot-based assays has been recommended to demonstrate LA activity, whereas solid-phase immunoassays allow the detection of anti-cardiolipin and anti-ß-2-glycoprotein-I antibodies. The diagnosis of APS requires the persistence of antiphospholipid antibodies for at least 12 weeks together with thrombotic and/or obstetric features of APS.


Assuntos
Trombofilia/diagnóstico , Anticoagulantes/uso terapêutico , Síndrome Antifosfolipídica/complicações , Síndrome Antifosfolipídica/diagnóstico , Deficiência de Antitrombina III/complicações , Deficiência de Antitrombina III/diagnóstico , Fator V/genética , Humanos , Inibidor de Coagulação do Lúpus/análise , Mutação de Sentido Incorreto , Mutação Puntual , Polimorfismo de Nucleotídeo Único , Deficiência de Proteína C/complicações , Deficiência de Proteína C/diagnóstico , Deficiência de Proteína S/complicações , Deficiência de Proteína S/diagnóstico , Protrombina/genética , Trombofilia/tratamento farmacológico , Trombofilia/etiologia , Trombofilia/genética
14.
BMC Med Genet ; 20(1): 2, 2019 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-30611230

RESUMO

BACKGROUND: Preeclampsia can lead to adverse maternal and perinatal outcomes. There are few studies on the association of preeclampsia with thrombophilia in Africa including Sudan. METHODS: A case -controls study was conducted at Saad Abualila Hospital in Khartoum, Sudan during the period of February through November 2017. The cases were women with preeclampsia and healthy pregnant women were the controls (180 women in each arm of the study). Genotyping for Factor-V Leiden 1691G/A and Prothrombin gene variation 20210G/A was done by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: There was no significant difference in the age, parity, body mass index (BMI) and the other characteristics between the cases and the controls. Genotypes distribution of Factor V Leiden 1691G/A and prothrombin gene 20210G/A in controls was in accordance with the Hardy-Weinberg equilibrium (P > 0.05). The factor V Leiden-variation was present in 9.6% of the cases compared with 0.6% of the controls, P < 0.001 (OR = 18.60, 95% CI = 2.38-136.1). Only 4 patients with severe preeclampsia had homozygous variation A/A and it was not detected in the controls. Prothrombin G20210A variations not detected neither in the cases nor in the controls group. CONCLUSIONS: High prevalence of Factor V Leiden 1691G/A variation in preeclamptic patients compared to controls suggest an involvement of this variation in predisposing to preeclampsia in this setting.


Assuntos
Fator V/genética , Polimorfismo de Nucleotídeo Único , Pré-Eclâmpsia/genética , Protrombina/genética , Adulto , Alelos , Índice de Massa Corporal , Estudos de Casos e Controles , Feminino , Genótipo , Heterozigoto , Homozigoto , Humanos , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Gravidez , Sudão , Adulto Jovem
15.
Eur J Med Genet ; 62(2): 93-95, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29879466

RESUMO

Loss-of-function mutations in coagulation cascade proteins lead to bleeding diasthesis. In contrast, gain-of-function mutations in these proteins, which are exceptionally rare, lead to hereditary thrombosis. This is best exemplified by Factor V (i.e., Factor V Leiden) and Factor II (i.e., p.Arg596Leu). Here, we report a family with hereditary thrombosis. The proposita presented with cerebral venous thrombosis accompanied by infarction at the age of 12 years. Despite anticoagulation therapy with oral warfarin, she later developed deep venous thrombosis in her hepatic portal veins at the age of 27 years. A medical exome analysis identified a de novo heterozygous mutation p.Tyr434His in Factor II, which was segregated within the family. A retrospective molecular diagnosis was made using a preserved surgical specimen from the proposita's mother, who had died 10 years earlier. The p.Tyr434 residue, which was substituted in the presently reported family, was located in "exosite I" of thrombin, a critical recognition site for fibrinogen, protein C, and thrombin aptamer. Therefore, the mutant thrombin likely exerted its thrombophilic effect by altering the affinity of thrombin to downstream substrates. Furthermore, the "exosite I" domain of thrombin was inhibited by the arthropod bleeding toxin Triabin (a protein discovered from the saliva of the blood-sucking triatomine bug Triatoma pallidipennis). Our observation that patients with a p.Tyr434His mutation exhibited recurrent thrombosis provides the first proof-of-concept in humans that a pharmacologic agent targeting "exosite I" could be an effective means of specifically modulating the thrombogenic-side of the coagulation system via the inhibition of factor II.


Assuntos
Proteínas de Insetos/farmacologia , Trombose Intracraniana/genética , Mutação de Sentido Incorreto , Protrombina/genética , Proteínas e Peptídeos Salivares/farmacologia , Trombose Venosa/genética , Adulto , Sítios de Ligação , Criança , Feminino , Heterozigoto , Humanos , Trombose Intracraniana/patologia , Masculino , Pessoa de Meia-Idade , Linhagem , Ligação Proteica , Protrombina/antagonistas & inibidores , Protrombina/química , Trombose Venosa/patologia
16.
J Thromb Thrombolysis ; 47(1): 87-95, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30284176

RESUMO

Laboratory testing for thrombophilia is complicated but essential for diagnosis. In 2017, the cobas® Factor II and Factor V Test (cobas F2F5 test) was launched for use with the cobas z 480 analyzer. This qualitative polymerase chain reaction test enables multiplex Factor II and Factor V testing with flexible reporting and workflow efficiency. Here, we report the results from studies investigating the performance of the cobas F2F5 test. Technical performance verification, clinical validation, external laboratory performance, and workflow comparison studies were performed. Fresh and frozen whole-blood and genomic DNA (gDNA) samples were tested, and several manual and automated DNA isolation methods were used. Bidirectional Sanger sequencing was used to verify genotypes identified by the cobas F2F5 test. One hundred percent agreement between the cobas F2F5 test and Sanger sequencing was observed for all genotypes. An external laboratory using remnant clinical samples also yielded 100% agreement between cobas F2F5 test results and their routine testing method. The cobas F2F5 test reduced the total sample processing time compared with the LightCycler® 1.2 platform (98.6 vs 420.2 min; 96 samples). Hemoglobin, extraction buffer, and ethanol contamination of the gDNA sample can lead to invalid results. The cobas F2F5 test has a high degree of accuracy for identification of Factor II and Factor V genotypes. This multiplex testing with short sample processing time can reduce handling errors and increase efficiency. Both manual and automated DNA isolation methods can be used with the cobas F2F5 test.


Assuntos
Testes de Coagulação Sanguínea/normas , Fator V/genética , Reação em Cadeia da Polimerase Multiplex/instrumentação , Mutação , Protrombina/genética , Trombofilia/genética , Testes de Coagulação Sanguínea/instrumentação , Técnicas de Laboratório Clínico/normas , Genótipo , Humanos , Reação em Cadeia da Polimerase Multiplex/normas , Análise de Sequência de DNA , Fatores de Tempo
17.
Clin Chim Acta ; 488: 221-225, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30439355

RESUMO

BACKGROUND: Practically, the initial step of genetic analysis is the extraction of DNA from blood or other cells, which is often time consuming and cost-intensive. We aimed at establishing a real-time PCR protocol for the detection of the venous thromboembolism associated mutations factor V Leiden (F5 c.1691G>A; p.R506Q) and prothrombin (F2) c.20210G>A from whole blood, without DNA extraction. METHODS: F5 c.1691G>A (p.R506Q) and F2 c.20210G>A mutations were determined in 205 EDTA anti-coagulated whole blood samples from patients who underwent routine clinical genotyping using the DirectBlood Genotyping PCR Kit (myPOLS Biotec, Konstanz, Germany) together with in-house developed TaqMan primer-probe assays. RESULTS: Validity score values of genotype calls using whole blood were similar and did not significantly differ compared to those using genomic DNA as substrate in PCR. Mutation analysis of 205 whole blood samples showed a negligible PCR dropout rate (one in 410 reactions) and were in 100% concordance with results obtained by conventional genotyping. CONCLUSION: We successfully established a robust and valid real-time PCR protocol for the detection of the venous thromboembolism associated mutations F5 c.1691G>A (p.R506Q) and F2 c.20210G>A directly from whole blood.


Assuntos
DNA/genética , Fator V/genética , Reação em Cadeia da Polimerase , Protrombina/genética , Tromboembolia Venosa/genética , DNA/isolamento & purificação , Genótipo , Humanos , Mutação , Tromboembolia Venosa/sangue
18.
J Mol Neurosci ; 67(2): 227-234, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30515700

RESUMO

Organophosphates (OP) are a major threat to the health of soldiers and civilians due to their use as chemical weapons in war and in terror attacks. Among the acute manifestations of OP poisoning, status epilepticus (SE) is bearing the highest potential for long-term damages. Current therapies do not prevent brain damage and seizure-related brain injuries in OP-exposed humans. Thrombin is a serine protease known to have a fundamental function in the clotting cascade. It is highly expressed in the brain where we have previously found that it regulates synaptic transmission and plasticity. In addition, we have found that an excess of thrombin in the brain leads to hyperexcitability and therefore seizures through a glutamate-dependent mechanism. In the current study, we carried out in vitro, ex vivo, and in vivo experiments in order to determine the role of thrombin and its receptor PAR-1 in paraoxon-induced SE. Elevated thrombin activity was found in the brain slices from mice that were treated (in vitro and in vivo) with paraoxon. Increased levels of PAR-1 and pERK proteins and decreased prothrombin mRNA were found in the brains of paraoxon-treated mice. Furthermore, ex vivo and in vivo electrophysiological experiments showed that exposure to paraoxon causes elevated electrical activity in CA1 and CA3 regions of the hippocampus. Moreover, a specific PAR-1 antagonist (SCH79797) reduced this activity. Altogether, these results reveal the importance of thrombin and PAR-1 in paraoxon poisoning. In addition, the results indicate that thrombin and PAR-1 may be a possible target for the treatment of paraoxon-induced status epilepticus.


Assuntos
Protrombina/metabolismo , Receptores de Trombina/metabolismo , Estado Epiléptico/metabolismo , Animais , Inibidores da Colinesterase/toxicidade , Hipocampo/metabolismo , Hipocampo/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Paraoxon/toxicidade , Protrombina/genética , Receptores de Trombina/agonistas , Receptores de Trombina/antagonistas & inibidores , Estado Epiléptico/etiologia
19.
Can J Cardiol ; 34(12): 1688.e13-1688.e15, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30527165

RESUMO

Transcatheter pulmonary valve (TPV) replacement is an effective therapy of right ventricular outflow tract conduit dysfunction. Acute complications after TPV implantation include infective endocarditis, stent fracture, and device dislocation. We present a novel, life-threatening complication: an acute, noninfectious TPV thrombosis. Within 24 hours after implantation of a Melody system (Medtronic, Inc, Minneapolis, MN), the patient developed an acute TPV thrombosis characterized by severe TPV stenosis on echocardiography and contrast filling defects on computed tomography pulmonary angiography images. Genetic testing revealed heterozygous prothrombin G20210A polymorphism and homozygous 4G/4G polymorphism of the plasminogen-activator-inhibitor. The patient recovered after surgical valve replacement with a pulmonary homograft.


Assuntos
Implante de Prótese de Valva Cardíaca , Próteses Valvulares Cardíacas/efeitos adversos , Estenose da Valva Pulmonar/etiologia , Valva Pulmonar/cirurgia , Trombose/etiologia , Angiografia por Tomografia Computadorizada , Heterozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Inativadores de Plasminogênio/genética , Polimorfismo Genético , Protrombina/genética , Estenose da Valva Pulmonar/diagnóstico por imagem , Trombose/diagnóstico por imagem
20.
Gac Med Mex ; 154(Supp 2): S15-S21, 2018.
Artigo em Espanhol | MEDLINE | ID: mdl-30532099

RESUMO

Objective: To examine the contribution the polymorphisms G20210A, G1691A and G10976A in the coagulation factors FII, FV, FVII, respectively; Glu298Asp and C677T in eNOS and 5,10 MTHFR in young Mexican population with cerebral infarction (CI). Methods: 224 patients ≤ 45 years of age with CI and 224 controls matched by age and gender were recruited from 2006 and 2014. The polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism. Results: We identified a significant difference in the genotype distribution of Glu298Asp (p = 0.001) and C677T (p = 0.01) polymorphisms between CI patients and control groups. The genotype distribution in the FII G20210A, FV G1691A and FVII G10976A polymorphisms were similar. There were independent factors for ischemic stroke: Glu298Asp and C677T polymorphisms, smoking; hypertension, and familial history of thrombotic disease. Conclusions: The Glu298Asp and C677T, but not FII G20210A, FV G1691A and FVII G10976A polymorphisms were associated with CI. Our results suggest that endothelial dysfunction and the synergist interaction with other factors such as smoking and hypertension contribute to CI in young individuals.


Assuntos
Infarto Cerebral/genética , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Óxido Nítrico Sintase Tipo III/genética , Acidente Vascular Cerebral/genética , Adulto , Isquemia Encefálica/genética , Fator V/genética , Fator VII/genética , Feminino , Genótipo , Humanos , Hipertensão/epidemiologia , Masculino , México , Reação em Cadeia da Polimerase , Polimorfismo Genético , Polimorfismo de Fragmento de Restrição , Protrombina/genética , Fumar/epidemiologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA