Relationship between serum vascular endothelial growth factor (VEGF) level and parameters of endothelial injury and/or dysfunction in patients with diabetes mellitus type 2 with or without microalbuminuria was investigated. Eighty-four diabetic patients were divided in two subgroups (42 each): normoalbuminuric (NAU) and microalbuminuric (MAU). Forty-two blood donors were in control group. Serum VEGF and plasma von Willebrand factor, soluble thrombomodulin, plasminogen activator inhibitor 1, thrombin-activatable fibrinolysis inhibitor (TAFI) and tissue plasminogen activator (t-PA) were measured using enzyme-linked immunosorbent assay in all subjects. VEGF was significantly higher in NAU compared to controls. The difference between MAU and controls was not statistically significant, but there was a trend toward significance. Only TAFI correlated with VEGF in MAU. An observed significant increase of serum VEGF level already in NAU suggests that serum VEGF could be a sensitive predictor of endothelial dysfunction in type 2 diabetes.

Disturbances of coagulation and fibrinolysis in type 2 diabetes mellitus (DM2) contribute to increased rates of macrovascular complications such as myocardial infarction and ischemic stroke. The aim of the study was to investigate the relationship among plasminogen activator inhibitor 1 (PAI-1), thrombin-activable fibrinolysis inhibitor (TAFI), tissue plasminogen activator (t-PA), prothrombin fragments 1+2 (F1+2), glycemic control, hypertension, sex and body mass index (BMI) in DM2 patients with normoalbuminuria and microalbuminuria. Forty-two normoalbuminuric (NAU), 42 microalbuminuric (MAU) patients with DM2 and 42 blood donors as control group were enrolled. TAFI, PAI-1, t-PA and F1+2 were assessed by enzyme-linked immunosorbent assay (ELISA) in all patients. TAFI was significantly increased in the MAU group, PAI-1 and F1+2 were increased in both groups and t-PA was not elevated in either group compared to controls. We found positive correlations in the NAU: TAFI and fibrinogen (r=0.65, P=0.02), PAI-1 and triglycerides (r=0.67, P=0.01), in the MAU: TAFI and F1+2 (r=0.48, P=0.02), TAFI and systolic blood pressure (r=0.53, P=0.01), PAI-1 and BMI (r=0.43, P<0.05). We found decreased fibrinolysis in DM2 patients presented with increased PAI-1 in both NAU and MAU. Hypofibrinolysis in MAU is further accented by the elevation of TAFI. TAFI-mediated inhibition of fibrinolysis in DM2 is regulated independently from PAI-1. Patient[Combining Acute Accent]s sex does not affect diabetes-related changes in hemostasis and fibrinolysis.

The GP6 gene encodes the GPVI, a crucial platelet membrane glycoprotein, for adequate platelet activation, adhesion and aggregation. The objectives of the present study were to assess the genetic variability of the GP6 gene in patients with platelet hyperaggregability phenotype, known as sticky platelet syndrome (SPS) manifesting as deep vein thrombosis (DVT), and/or pulmonary embolism, and in controls; and to evaluate its role in the pathogenesis of venous thromboembolism (VTE) in SPS. Seventy-seven patients with SPS and 77 healthy blood donors as controls were enrolled. Light transmission aggregometry was used to diagnose SPS according to the method of Mammen and Bick. Seven single-nucleotide polymorphisms (SNPs) of the GP6 gene (rs1654410, rs1671153, rs1654419, rs11669150, rs12610286, rs1654431, rs1613662) were assessed using restriction fragment length polymorphism analysis. A significant association between 1613662-G [P < 0.05, odds ratio (OR) 2.087, confidence interval (CI) 1.049-4.148], 1654419-A (P < 0.05, OR 2.161, CI 1.020-4.577) and VTE was found in patients with SPS. The analysis based on SPS type revealed a significantly higher occurrence of 1671153-G (P < 0.05, OR 2.317, CI 1.103-4.865) and 1654419-A (P < 0.05, OR 2.317, CI 1.103-4.865) in the SPS type II compared to the control group. No association between the studied GP6 genotypes and the severity of VTE (pulmonary embolism vs. DVT) was found. In the patients, significant positive relationship between the 1671153-G, 1654419-A, 1613662-G alleles and male sex was observed. GP6 SNPs 1613662-G, 1671153-G and 1654419-A alleles are associated with an increased risk of VTE in SPS. They could contribute to the SPS phenotype.

No abstract

The exact pathogenesis of SPS is not sufficiently explained. Our findings seem to support the idea that SPS might have an autosomal dominant hereditary fashion.

Background: Rotational thrombelastometry (ROTEM) is a real-time clotting test that provides insight into clotting factors, the fibrinolytic system and platelet function. We obtained the longitudinal values on ROTEM in normal pregnancy and in puerperium. Material and Methods: After ethics committee approval and subject informed consent, citrated blood was sampled from healthy pregnants four times during pregnancy and one time postpartum. As controls we used nonpregnant women undergoing voluntary blood donation. Extem and Intem tests and basic coagulation test were carried out. Results: We included 112 women in our study, 55 non-pregnant women (controls) and 57 healthy pregnants with 5 samplings. The values of maximum clot firmness (MCF - in EXTEM and INTEM) were significantly higher up to 34th-36th week of pregnancy than those in non-pregnant subjects. MCF in 6th-7th week after delivery was significantly higher in both tests. Clotting time (CT) in pregnant women was significantly shorter (EXTEM) compared to non-pregnant subjects. We also found a very strong correlation between MCF and platelet count in all gestational weeks.' Conclusions: Rotation thromboelastometry clearly demonstrates the hypercoagulability in pregnancy and can reflect the higher risk of venous thromboembolism in both pregnancy and puerperium. Strong correlation between MCF and platelet count can suggest role of platelets in hypercoagulability in pregnant women. This study provides a better knowledge about physiological changes in ROTEM measurement during pregnancy and postpartum.

Bortezomib (PS-341, or Velcade), reversible inhibitor of 20S proteasome approved for the treatment of multiple myeloma and mantle cell lymphoma, exhibited a cytotoxic effect toward other malignancies including leukaemia. In this study, we have documented that incubation of both HL-60 and K562 leukaemia cells with nanomolar concentrations of bortezomib is associated with the death of HL-60 cells observed within 24 hours of incubation with bortezomib and the death of K562 cells that were observed after 72 hours of incubation with bortezomib. The relative resistance of K562 cells to bortezomib correlated well with significantly higher expression of HSP27, HSP70, HSP90α, HSP90β and GRP75 in these cells. Incubation of both HL-60 and K562 cells with bortezomib induced a cleavage of HSP90β as well as expression of HSP70 and HSP90β but bortezomib did not affect levels of HSP27, HSP90α, GRP75 and GRP78. The death of both types of cells was accompanied with proteolytic activation of caspase 3 that was observed in HL-60 cells and proteolytic degradation of procaspase 3 in K562 cells. Our study has also pointed to essential role of caspase 8 in bortezomib-induced cleavage of HSP90β in both HL-60 and K562 cells. Finally, we have shown that bortezomib induced activation of caspase 9/caspase 3 axis in HL-60 cells, while the mechanism of death of K562 cells remains unknown.

The mechanisms of fibrinolysis have been suggested to be linked to the pathogenesis of peripheral artery disease. The impact of therapeutic angiogenesis on the parameters of fibrinolysis was studied in critical limb ischemia (CLI). CLI patients (N = 29) and blood donors as controls (N = 29) were enrolled. Bone marrow (600 ± 50 ml) was centrifuged (3200g, 20 min, 22°C), bone marrow-derived mononuclear cells (100-120 ml) were separated by Optipress I and implanted into the ischemic limb using intramuscular injections. ELISA was employed for the assessment of plasma tissue plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) levels. Patients were followed-up prior to the procedure and after 1, 3 and 6 months. All stage-IV patients (N = 22) had ischemic lesions. The lesions resolved in 10 patients. Five patients underwent major amputation; they all were stage-IV. Ischemic lesions persisted in seven patients beyond 6 months. The t-PA levels were higher in patients compared with the healthy controls both at baseline (P < 0.01) and after 6 months (P < 0.05). No significant changes were observed in the t-PA levels during the follow-up. PAI-1 was higher in patients than in the healthy individuals at baseline (P < 0.001) and at month 1 (P < 0.05). However, no difference in PAI-1 levels between the patients and the healthy individuals was found after 3 and 6 months. The PAI-1 levels were significantly downregulated during the follow-up compared with the baseline (P < 0.0001). Therapeutic angiogenesis for the CLI downregulates PAI-1 levels, thus having a systemic effect on fibrinolysis.