Factors influencing quality of anticoagulation control and warfarin dosage in patients after aortic valve replacement within the 3 months of follow up.

Warfarin dosage estimation using the pharmacogenetic algorithms has been shown to improve the quality of anticoagulation control in patients with atrial fibrillation. We sought to assess the genetic, demographic and clinical factors that determine the quality of anticoagulation in patients following aortic valve replacement (AVR). We studied 200 consecutive patients (130 men) aged 63 ± 12.3 years, undergoing AVR, in whom warfarin dose was established using a pharmacogenetic algorithm. The quality of anticoagulation within the first 3 months since surgery was expressed as the time of international normalized ratio (INR) in the therapeutic range (TTR). The median TTR in the entire cohort was 59.6% (interquartile range, 38.7 - 82.7). Ninety-nine (49.5%) patients with TTR ≥ 60% did not differ from those with poor anticoagulation control (TTR < 60%) with regard to demographic and cardiovascular risk factors. Coronary artery disease (n = 84, 42%) and previous stroke (n = 5, 2.5%) predicted higher TTR, while possession of CYP2C9*2 variant allele (n = 49, 25%) was associated with lower TTR (P = 0.01). In turn, VKORC1 c.-1639A, CYP2C9*2 and *3 variants were independently associated with actual warfarin dose (P < 0.0001). In AVR patients better anticoagulation control is observed in patients with coronary artery disease and history of stroke, which might result in part from previous lifestyle modification and therapy. Possession of CYP2C9*2 and/or CYP2C9*3 allele variants is associated with lower TTR values and warfarin dose variations in AVR patients, the latter affected also by VKORC1 c.-1693G>A polymorphism.

Hypoglycosylation is a common finding in antithrombin deficiency in the absence of a SERPINC1 gene defect.

UNLABELLED: Essentials We investigated the molecular base of antithrombin deficiency in cases without SERPINC1 defects. 27% of cases presented hypoglycosylation, transient in 62% and not restricted to antithrombin. Variations in genes involved in N-glycosylation underline this phenotype. These results support a new form of thrombophilia. Click here to listen to Dr Huntington's perspective on thrombin inhibition by the serpins SUMMARY: Background Since the discovery of antithrombin deficiency, 50 years ago, few new thrombophilic defects have been identified, all with weaker risk of thrombosis than antithrombin deficiency. Objective To identify new thrombophilic mechanisms. Patients/methods We studied 30 patients with antithrombin deficiency but no defects in the gene encoding this key anticoagulant (SERPINC1). Results A high proportion of these patients (8/30: 27%) had increased hypoglycosylated forms of antithrombin. All N-glycoproteins tested in these patients (α1-antitrypsin, FXI and transferrin) had electrophoretic, HPLC and Q-TOF patterns indistinguishable from those of the congenital disorders of glycosylation (rare recessive multisystem disorders). However, all except one had no mental disability. Moreover, intermittent antithrombin deficiency and hypoglycosylation was recorded in five out of these eight patients, all associated with moderate alcohol intake. Genetic analysis, including whole exome sequencing, revealed mutations in different genes involved in the N-glycosylation pathway. Conclusions Our study provides substantial and novel mechanistic insights into two disease processes, with potential implications for diagnosis and clinical care. An aberrant N-glycosylation causing a recessive or transient antithrombin deficiency is a new form of thrombophilia. Our data suggest that congenital disorders of glycosylation are probably underestimated, especially in cases with thrombosis as the main or only clinical manifestation.

Reduced plasma fibrin clot permeability and susceptibility to lysis are associated with increased risk of postthrombotic syndrome.

BACKGROUND: The postthrombotic syndrome (PTS) is a severe complication of deep vein thrombosis (DVT). Reduced plasma clot permeability and lysability have been linked to DVT and residual vein obstruction. OBJECTIVES We investigated whether altered fibrin clot properties are associated with the occurrence of PTS. PATIENTS AND METHODS: Plasma fibrin clot permeability (Ks ) and lysability were investigated in a cohort of 197 consecutive patients aged 18 to 65 years recruited 3 months following the first-ever DVT. Patients with severe thrombophilia or comorbidities known to adversely affect clot phenotype were ineligible. RESULTS: During a 1-year follow-up PTS developed in 48 (24%) patients, who were characterized by lower Ks , prolonged fibrin clot lysis time (CLT) and slower release of D-dimer from clots (D-Drate ), together with higher plasma D-dimer, C-reactive protein and thrombin-activatable fibrinolysis inhibitor (TAFI). No PTS-associated differences in fibrinogen, thrombin generation, factor VIII, other fibrinolysis proteins and the quality of anticoagulation were observed. Ks (r = -0.71), CLT (r = 0.45), D-Drate (r = -0.30) and TAFI activity (r = 0.38) were associated with the Villalta scale (all P < 0.05). Recurrent VTE occurred also more commonly in PTS patients during follow-up and the 26 (13.2%) patients had lower Ks , longer CLT and lower D-Drate (all P < 0.05). A multivariate model adjusted for age, body mass index, fibrinogen and glucose showed that independent predictors of PTS were idiopathic DVT, plasma D-dimer, Ks , D-Drate , tissue plasminogen activator and TAFI activity. CONCLUSIONS: This study demonstrates that formation of more compact fibrin clots displaying impaired susceptibility to lysis predisposes to PTS.

Impaired fibrinolysis is associated with the severity of aortic stenosis in humans.

BACKGROUND: A role of fibrinolysis in the pathogenesis of aortic valve stenosis (AS) is unknown, although fibrinolytic proteins have been detected in aortic stenotic valves. OBJECTIVE: To investigate whether impaired fibrinolysis could be associated with AS. METHODS AND RESULTS: We studied 74 patients with AS (43 male, 31 female, aged 62.7 ± 10.7 years) without documented atherosclerotic valvular disease scheduled for isolated valve replacement and 68 controls. The plasma fibrin clot lysis time (CLT) in the presence of tissue factor (TF) and tissue plasminogen activator (tPA), along with plasma plasminogen activator inhibitor-1 (PAI-1) were determined. Valvular expression of fibrin and PAI-1 together with macrophages and mast cells (MC) was evaluated by immunostaining. Patients with AS compared with controls were characterized by a prolonged CLT (median, 110 [54-153] vs. 92.5 [58-115] min, P = 0.0007) and increased plasma PAI-1 (78.6 [35.5-149] vs. 38.5 [18-61] ng mL(-1) , P < 0.0001). CLT was correlated with maximal (r = 0.43, P = 0.0002) and mean (r = 0.38, P = 0.001) transvalvular pressure gradients, and aortic valve area (r = -0.59, P < 0.0001). In AS patients, the CLT was positively correlated with the valve leaflet thickness (r = 0.67, P = 0.003), the degree of valve calcification (r = 0.65, P < 0.00001), valvular fibrin (r = 0.54, P = 0.007) and PAI-1 expression (r = 0.48, P = 0.007). Double-immunostaining revealed colocalization of valvular PAI-1 with MC (87 ± 17% cells) and macrophages (48 ± 11% cells) within stenotic valves. CONCLUSIONS: Hypofibrinolysis might be a marker of severe AS and be implicated in AS progression.

Mast cells in human stenotic aortic valves are associated with the severity of stenosis.

Aortic valve stenosis (AS) is characterized by extensive calcification of the aortic valve leaflets and infiltration of inflammatory cells. Activated mast cells (MCs) may participate in the induction of fibrosis and calcification with ensuing valve stiffening. We sought to investigate whether the number of MCs within stenotic aortic valves is associated with the severity of AS. We studied 43 patients (19 men, 24 women) with dominant AS (age, 64.2 ± 5.9 years; mean transvalvular pressure gradient, 62.11 ± 24.47 mmHg) without atherosclerotic vascular disease, undergoing elective aortic valve replacement. MCs were detected in the excised valves by immunostaining. Aortic valves from five healthy subjects obtained on autopsy served as negative controls. The number of tryptase- and chymase-positive MCs was increased in AS valves compared with the control valves (6.9 [2.3-18.9]/mm(2) vs. 0.7 [0-2.2]/mm(2), P = 0.0001 and 3.2 [2.1-9.4]/mm(2) vs. 0.3 [0-1.9]/mm(2), P = 0.002, respectively). MCs that colocalized with macrophages and neovessels were detected mainly in the calcified regions of the leaflets. The number of MCs positively correlated with maximal (r = 0.73, P < 0.0001) and mean (r = 0.78, P < 0.0001) gradients and maximal aortic jet velocity (r = 0.68, P = 0.0005). An inverse correlation with aortic valve area (r = -0.71, P = 0.0001) was also observed. Multivariate regression analysis revealed that MC number and valve thickness were significantly associated with mean transvalvular gradient (R (2) = 0.74, P < 0.000001) in AS patients. Increased MC number within human stenotic aortic valves is associated with the severity of AS.

The increased plasma C-reactive protein and interleukin-6 levels in patients undergoing coronary artery bypass grafting surgery are associated with the interleukin-6-174G > C gene polymorphism.

BACKGROUND: The interleukin-6 (IL-6) promoter -174G/C polymorphism (rs1800795) is associated with enhanced systemic inflammatory response to injury. However, data on the effect of this polymorphism on inflammatory markers in patients undergoing coronary artery bypass grafting surgery (CABG) are inconsistent. The aim of our study was to investigate whether -174G/C IL-6 polymorphism affects plasma IL-6 and C-reactive protein (CRP) concentrations in patients undergoing CABG. METHODS: A total of 179 consecutive white patients (77% men, aged 65 +/- 8.6 standard deviation [SD] y) scheduled for elective isolated CABG were studied. Pre- and postoperative CRP and IL-6 levels were analysed in relation to the 174G/C IL-6 polymorphism determined by using TaqMan single-nucleotide polymorphism genotyping technique. RESULTS: The genotype distribution was as follows: GG -46 (26%), GC -93 (52%) and CC -40 (22%). The C allele carriers had higher baseline CRP (4.1 +/- 0.35 versus 2.4 +/- 0.59 mg/L, P = 0.02) and IL-6 levels (3.0 +/- 0.17 versus 2.2 +/- 0.3 pg/mL, P = 0.02) than GG patients. Five to seven days after CABG, CRP levels rose by 54% (P = 0.03), and IL-6 levels tended to be higher (P = 0.07) in -174C allele carriers than the non-carriers. There were no associations between -174G/C IL-6 polymorphism and any demographic-, clinical- or procedure-related variables as well as major adverse cardiovascular events. Multivariate regression analysis, including sex, age, body mass index, hypercholesterolaemia, smoking, hypertension diabetes, identified CG + CC genotype as the only independent predictor of preoperative CRP and IL-6 levels. CONCLUSIONS: The presence of the -174C allele determines to some extent higher plasma CRP and IL-6 concentrations pre- and postoperatively in CABG patients.

Biomechanical signals exert sustained attenuation of proinflammatory gene induction in articular chondrocytes.

OBJECTIVES: Physical therapies are commonly used for limiting joint inflammation. To gain insight into their mechanisms of actions for optimal usage, we examined persistence of mechanical signals generated by cyclic tensile strain (CTS) in chondrocytes, in vitro. We hypothesized that mechanical signals induce anti-inflammatory and anabolic responses that are sustained over extended periods. METHODS: Articular chondrocytes obtained from rats were subjected to CTS for various time intervals followed by a period of rest, in the presence of interleukin-1beta (IL-1beta). The induction for cyclooxygenase (COX-2), inducible nitric oxide synthase (iNOS), matrix metalloproteinase (MMP)-9, MMP-13 and aggrecan was analyzed by real-time polymerase chain reaction (PCR), Western blot analysis and immunofluorescence. RESULTS: Exposure of chondrocytes to constant CTS (3% CTS at 0.25 Hz) for 4-24 h blocked more than 90% (P<0.05) of the IL-1beta-induced transcriptional activation of proinflammatory genes, like iNOS, COX-2, MMP-9 and MMP-13, and abrogated inhibition of aggrecan synthesis. CTS exposure for 4, 8, 12, 16, or 20 h followed by a rest for 20, 16, 12, 8 or 4h, respectively, revealed that 8h of CTS optimally blocked (P<0.05) IL-1beta-induced proinflammatory gene induction for ensuing 16 h. However, CTS for 8h was not sufficient to inhibit iNOS expression for ensuing 28 or 40 h. CONCLUSIONS: Data suggest that constant application of CTS blocks IL-1beta-induced proinflammatory genes at transcriptional level. The signals generated by CTS are sustained after its removal, and their persistence depends upon the length of CTS exposure. Furthermore, the sustained effects of mechanical signals are also reflected in their ability to induce aggrecan synthesis. These findings, once extrapolated to human chondrocytes, may provide insight in obtaining optimal sustained effects of physical therapies in the management of arthritic joints.

Opposite effects of mast cell degranulation by compound 48/80 on peritoneal inflammation in Swiss and CBA mice.

The murine strains differ in the number of peritoneal mast cells. Degranulation of peritoneal mast cells by single injection of compound 48/80 (1.2 mg/kg) followed by zymosan-induced (2 mg/ml, 0.5 ml/mouse) peritoneal inflammation caused either inhibition or enhancement of an early influx (at 4 h of peritonitis) of exudatory leukocytes in Swiss and CBA mice, respectively. These opposite effects correspond with statistically significant differences in the number of peritoneal mast cells in the intact Swiss (11 x 10(3)) and CBA (39 x 10(3)) mice.