Diurnal Oscillations of Fibrinolytic Parameters in Patients with Acute Myocardial Infarction and Their Relation to Platelet Reactivity: Preliminary Insights.

There is limited information about diurnal changes in fibrinolysis parameters after acute myocardial infarction (AMI) and their relationship with on-treatment platelet reactivity. The aim of this study was to assess tissue plasminogen activator (t-PA), plasminogen activator inhibitor type-1 (PAI-1), α2-antiplasmin (α2-AP) activity, and plasmin-antiplasmin (PAP) complexes in 30 AMI patients taking dual antiplatelet therapy (DAPT), i.e., acetylsalicylic acid and clopidogrel. Fibrinolytic parameters were assessed at four time points (6 a.m., 10 a.m., 2 p.m., and 7 p.m.) on the third day after AMI using immunoenzymatic methods. Moreover, platelet reactivity was measured using multiple-electrode aggregometry, to assess potential differences in fibrinolytic parameters in low/high on-aspirin platelet reactivity and low/high on-clopidogrel platelet reactivity subgroups of patients. We detected significant diurnal oscillations in t-PA and PAI-1 levels in the whole study group. However, PAP complexes and α2-AP activity were similar at the analyzed time points. Our study reveals a potential impact of DAPT on the time course of fibrinolytic parameters, especially regarding clopidogrel. We suggest the presence of diurnal variations in t-PA and PAI-1 concentrations in AMI patients, with the highest levels midmorning, regardless of platelet reactivity. Significantly elevated levels of PAI-1 during the evening hours in clopidogrel-resistant patients may increase the risk of thrombosis.

Adipokine levels and carbohydrate metabolism in patients diagnosed de novo with polycystic ovary syndrome.

INTRODUCTION: Central obesity appears to play a major role in the pathogenesis of metabolic disorders in polycystic ovary syndrome. Insulin resistance and carbohydrate disorders are associated with dysfunctional secretion of various adipokines by the adipose tissue. OBJECTIVES: This study aimed to evaluate leptin, apelin, and visfatin against a background of carbohydrate metabolism parameters in patients diagnosed de novo with polycystic ovary syndrome (PCOS). MATERIAL AND METHODS: The study group consisted of 40 patients with PCOS (mean age, 29 years) diagnosed in accordance with the American Society for Reproductive Medicine criteria from 2003. The control group consisted of 37 clinically healthy women (mean age, 26 years). All controls had regular menses and no clinical or biochemical signs of hyperandrogenism. Concentrations of leptin, apelin, visfatin, and insulin were measured by immunoenzymatic methods. Glucose concentrations were determined using spectrophotometry. RESULTS: Significantly higher concentrations of leptin, insulin, homeostatic model assessment for insulin resistance (HOMA-IR) index, and the immunoreactive insulin (IRI)/glucose index were found in the PCOS group than in the control group. Notably, the concentration of apelin was over five times lower in the PCOS group than in the control group. In patients with PCOS, a positive correlation was found between the concentrations of insulin and leptin and concentrations of leptin and IRI/glucose. Patients of the PCOS group with body mass index (BMI) ≥  25 had significantly higher values of leptin, insulin, HOMA-IR index, and IRI/glucose index than patients of the PCOS group with normal BMI. In the PCOS group, a positive correlation was found between BMI and leptin concentration (r = 0.7176; p < 0.0001) and carbohydrate metabolism, such as insulin (r = 0.5524; p = 0.0003), glucose (r = 0.3843; p = 0.0157), HOMA-IR (r = 0.5895; p < 0.0001), and IRI/glucose (r = 0.3872; p = 0.0163). These findings were not observed in the control group. CONCLUSIONS: (1) Increased leptin concentration observed in women diagnosed de novo with PCOS as well as positive correlations between leptin and HOMA-IR, and IRI/glucose and BMI may indicate a potential role of leptin in the reduction of tissue sensitivity to insulin. (2) Significantly lower apelin concentration in the PCOS group (>5 fold) than in the control group, associated with a concomitant increase in leptin, may also contribute to carbohydrate metabolism disorders occurring in the course of PCOS.

Assessment of endothelial progenitor cells, VEGF-A and SDF-1α in Hodgkin's lymphoma.

Recently, there is great interest in vasculogenesis, a process of the formation of new blood vessels from progenitor cells or angioblasts, in the pathogenesis of cancer. To the best of our knowledge, the evaluation of endothelial progenitor cells (EPCs) in Hodgkin's lymphoma has not yet been reported. The aim of the present study was to assess the number of EPCs and selected cytokines, such as vascular endothelial growth factor (VEGF-A) and stromal cell-derived factor (SDF-1α) involved in vasculogenesis in Hodgkin's lymphoma patients. The study was conducted in a group of 42 patients with Hodgkin's lymphoma (eight patients with relapsed Hodgkin's lymphoma and 34 patients before the first treatment) and 30 healthy controls. The number of EPCs defined as CD31(+), CD34(+), CD45(-), CD133(+) was analysed on FacsCalibur flow cytometer and the concentration of VEGF-A and SDF-1α was assessed by ELISA. The study showed that there was a significantly higher EPCs number and VEGF-A concentration in the blood of Hodgkin's lymphoma patients compared to healthy individuals (8.20 vs. 0.55 cells/µl; P < 0.000001; 85.10 vs. 25.33 pg/ml, P = 0.000017; respectively). Detailed analysis revealed that there was elevated EPCs number in both study subgroups as compared to the control group. However, there was no difference in VEGF concentration between recurrent Hodgkin's lymphoma patients and the control group. A significant positive correlation was found between the number of EPCs and VEGF-A concentration (R = 0.31, P = 0.047). Significantly higher EPCs number combined with increased VEGF-A concentration, found in Hodgkin's lymphoma patients before the first treatment, suggest stimulation of new blood vessels formation, which may in turn contribute to tumour growth and metastasis in these patients.

Increased number of endothelial progenitor cells in the blood of patients with psoriasis vulgaris.

Introduction: Psoriasis vulgaris (PsV) is a common dermatosis characterized by excessive activation of neovascularization. Latest research has shown that endothelial progenitor cells (EPCs) are a crucial factor involved in the repair of endothelial injury and formation of new blood vessels, in a process termed postnatal vasculogenesis. However, the exact mechanism of creating psoriatic skin patches and the involvement of EPCs in this process remains unknown. Aim: To evaluate the number of EPCs in the blood of patients with PsV, characterized by the expression of specific cell surface markers, including CD45-, CD31+, CD34+ and CD133+. Material and methods: A total of 49 patients suffering from PsV and 40 healthy volunteers were enrolled in the study. The number of EPCs in each of the volunteers' whole blood samples was measured with a FACSCalibur flow cytometer using monoclonal antibodies directed against antigens specific for EPCs. Results: The number of EPCs was significantly higher in patients with psoriasis compared with the controls (p = 0.0007) and inversely correlated with disease severity assessed by PASI score (R = -0.2935, p = 0.0407). Statistical analysis did not show significant relations between the count of EPCs and age, body mass index, gender, disease duration, blood pressure, extent of itching, severity and frequency of pruritus, presence of bruises, vitamin D supplementation and smoking habit. Conclusions: The results of our studies indicate that patients with psoriasis showed an increased mobilization of EPCs compared with healthy individuals which correlated negatively with disease severity.

Endothelial Progenitor Cells as a Marker of Vascular Damage But not a Predictor in Acute Microangiopathy-Associated Stroke.

BACKGROUND: The aim of the study was to assess the number of endothelial progenitor cells (EPCs) in patients with acute stroke due to cerebral microangiopathy and evaluate whether there is a relationship between their number and clinical status, radiological findings, risk factors, selected biochemical parameters, and prognosis, both in ischemic and hemorrhagic stroke. METHODS: In total, 66 patients with lacunar ischemic stroke, 38 patients with typical location hemorrhagic stroke, and 22 subjects from the control group without acute cerebrovascular incidents were included in the prospective observational study. The number of EPCs was determined in serum on the first and eighth day after stroke onset using flow cytometry and identified with the immune-phenotype classification determinant (CD)45-, CD34+, CD133+. RESULTS: We demonstrated a significantly higher number of EPCs on the first day of stroke compared to the control group (med. 17.75 cells/µL (0-488 cells/µL) vs. 5.24 cells/µL (0-95 cells/µL); p = 0.0006). We did not find a relationship between the number of EPCs in the acute phase of stroke and the biochemical parameters, vascular risk factors, or clinical condition. In females, the higher number of EPCs on the first day of stroke is related to a favorable functional outcome on the eighth day after the stroke onset compared to males (p = 0.0355). We found that a higher volume of the hemorrhagic focus on the first day was correlated with a lower number of EPCs on the first day (correlation coefficient (R) = -0.3378, p = 0.0471), and a higher number of EPCs on the first day of the hemorrhagic stroke was correlated with a lower degree of regression of the hemorrhagic focus (R = -0.3896, p = 0.0367). CONCLUSION: The study showed that endothelial progenitor cells are an early marker in acute microangiopathy-associated stroke regardless of etiology and may affect the radiological findings in hemorrhagic stroke. Nevertheless, their prognostic value remains doubtful in stroke patients.

Inhibitors of the blood coagulation process in patients with essential thrombocythemia.

: The aim of the study was to assess the activity of protein C, protein S and tissue factor pathway inhibitor in relation to the risk factors for thrombotic complications in patients with essential thrombocythemia.The study group consisted of 45 newly diagnosed patients with essential thrombocythemia. Protein S activity was determined by chromogenic method. Activities of protein C and tissue factor pathway inhibitor (TFPI) were determined using ELISAs.Significantly lower protein C and protein S activity but higher TFPI activity were found in patients with ET in comparison with the control group. TFPI activity was higher in women as compared to men, and in patients over 60 years of age compared with patients below 60 years of age. TFPI activity was higher in patients with leukocytes count at least 11 g/l than in patients with leukocytes count below 11 g/l and the difference almost reached statistical significance. Significantly lower protein C activity was found in patients with the JAK2V617F mutation, in comparison with essential thrombocythemia patients JAK2V617F (-).The reduced protein C and protein S activity may be one of the pathogenic factors of increased prothrombotic state in essential thrombocythemia patients. The decreased protein C activity in patients with the JAK2 V617F mutation seems to confirm the significant role of this mutation in the pathogenesis of thrombotic complications in essential thrombocythemia patients. Significantly increased TFPI activity in essential thrombocythemia patients above 60 years of age and with leukocyte count above 11 g/l expresses the activation of the compensatory mechanism for increased prothrombotic activity.

Diurnal variations in tissue factor and tissue factor pathway inhibitor concentrations in relation to on-treatment platelet reactivity: an analysis of patients with acute myocardial infarction.

The aim of the study was to evaluate diurnal changes of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) concentrations in relation to on-treatment platelet reactivity. The study group included 51 patients with acute myocardial infarction (AMI) treated with primary percutaneous coronary intervention and dual antiplatelet therapy. TF and TFPI concentrations were assessed using enzyme-linked immunosorbent assay kits. We found a significant increase of TF concentration in clopidogrel-resistant, but not clopidogrel-sensitive, patients at 10.00 a.m. (410.66 pg/mL) in comparison with 6.00 a.m. (250.99 pg/mL), 14.00 p.m. (255.12 pg/mL) and 19.00 p.m. (267.58 pg/mL). Moreover, TF concentration at 10.00 a.m. was 30% higher in clopidogrel-resistant than clopidogrel-sensitive patients (p = .043). We failed to demonstrate diurnal variation in TFPI concentration in clopidogrel-resistant patients. However, TFPI concentration in clopidogrel-sensitive patients was significantly higher at 10.00 a.m. as compared with other sampling points (p < .05). We observed a marked elevation in TF concentration at 10.00 a.m. only in aspirin-resistant patients and a significant increase in TFPI concentration at 10 a.m. only in aspirin-sensitive patients. Our findings suggest the presence of diurnal variations in TF and TFPI concentrations in AMI patients, with the highest thrombotic risk in patients with high on-treatment platelet reactivity in the midmorning.

Activation of TF-Dependent Blood Coagulation Pathway and VEGF-A in Patients with Essential Thrombocythemia.

Background and objectives: Recent studies suggest that a vascular endothelial growth factor (VEGF-A) may be involved in the thrombotic process by stimulating the expression of tissue factor in vascular endothelial cells. Tissue factor (TF) can also stimulate the transcription of the gene encoding VEGF-A. The relationship between coagulation and angiogenesis in myeloproliferative neoplasms is not fully understood. The aim of this study was to evaluate the concentration of TF in relation to VEGF-A in the blood of patients with essential thrombocythemia (ET). Patients and methods: The study group consisted of 130, newly diagnosed patients with ET (mean age 61 years). The control group consisted of 35 healthy volunteers (mean age 51 years). Concentrations of VEGF-A, TF, and tissue factor pathway inhibitor (TFPI) were analysed using immunoenzymatic methods. TF and TFPI activities were performed using chromogenic assays. Results: The median concentration of TF Ag was 3-fold higher and the TF activity was more than 15-fold higher in ET patients than in normal individuals. There were no statistically significant differences in the TFPI concentration and activity between groups. VEGF-A was significantly increased in patients with ET (p < 0.000001). Analysis of correlations revealed a positive correlation between VEGF-A and TF Ag as well as a positive correlation between VEGF-A and TFPI activity. Conclusions: The simultaneous increase of TF concentration and activity, VEGF-A in the blood of patients with ET, as well as a positive correlation between the concentration of TF and VEGF-A demonstrates the coexistence of TF-dependent coagulation and activation of angiogenesis.

Selected Parameters of Angiogenesis and the JAK2, CALR, and MPL Mutations in Patients With Essential Thrombocythemia.

The aim of the study was to evaluate selected angiogenic factors in patients with essential thrombocythemia (ET) depending on JAK2V617F, calreticulin gene (CALR) and myeloproliferative leukemia virus oncogene (MPL) mutations. Sixty ET patients and 20 healthy volunteers were enrolled in the study. The following tests were performed: vascular endothelial growth factor- A (VEGF-A), soluble vascular endothelial growth factor receptor-1 (sVEGFR-1),soluble vascular endothelial growth factor receptor-2 (sVEGFR-2), platelet-derived growth factor( PDGF-BB), and stromal-derived factor-1α (SDF-1α). We observed an increased PDGF-BB level in patients with ET compared to the controls. Patients with CALR mutation had significantly higher concentration of PDGF-BB and lower concentration of SDF-1α than patients with JAK2V617F mutation. High concentration of PDGF-BB and low concentration of SDF-1α in patients with CALR(+) ET may indicate a contribution of these chemokines in disturbed Ca2+ metabolism in platelets.

VEGF-A, sVEGFR-1, and sVEGFR-2 in BCR-ABL negative myeloproliferative neoplasms.

BACKGROUND AND OBJECTIVE: Data from the literature indicate the relationship between the bone marrow microvessel density and the blood parameters of angiogenesis. The aim of this study was to evaluate selected parameters of angiogenesis (VEGF-A, sVEGFR-1, and sVEGFR-2) and their correlations with white blood cells, platelets, and red blood cells. MATERIALS AND METHODS: The study included 72 patients (mean age, 61.84 years) with myeloproliferative neoplasms (MPNs): essential thrombocythemia (ET) (n=46), polycythemia vera (PV) (n=19), and primary myelofibrosis (PMF) (n=7). Serum VEGF-A, sVEGFR-1, and sVEGFR-2 were determined using the ELISA assay. RESULTS: We observed a significantly higher level of VEGF-A and reduced concentrations of sVEGFR-1 and sVEGFR-2 in the whole group of patients with MPNs as compared to controls. Detailed analysis confirmed significantly higher level of VEGF-A and lower concentration of sVEGFR-2 in each subgroups of MPNs patients. However, sVEGFR-1 concentrations were significantly lower only in PV and ET patients. CONCLUSIONS: The study showed an increased level of VEGF-A, which may indicate the intensity of neoangiogenesis in the bone marrow. Decreased sVEGFR-1 and sVEGFR-2 in the blood of patients with MPNs may reflect consumption of these soluble receptors.

Which platelet function test best reflects the in vivo plasma concentrations of ticagrelor and its active metabolite? The HARMONIC study.

Aim of this study was assessment of the relationship between concentrations of ticagrelor and its active metabolite (AR-C124910XX) and results of selected platelet function tests. In a single-centre, cohort study, patients with myocardial infarction underwent blood sampling following a 180 mg ticagrelor loading dose intake (predose, 1, 2, 3, 4, 6, 12, 24 hours postdose) to perform pharmacokinetic and pharmacodynamic assessments. Platelet reactivity was evaluated using the VASP-assay, the VerifyNow device and the Multiplate analyzer. Analysis of 36 patients revealed high negative correlations between ticagrelor concentrations and platelet reactivity evaluated with all three platelet function tests (the VASP-assay: RS=-0.722; p<0.0001; the VerifyNow device: RS=-0.715; p<0.0001; the Multiplate analyzer: RS=-0.722; p<0.0001), with no significant differences between correlation coefficients. Similar results were found for AR-C124910XX. Platelet reactivity values assessed with all three methods generally correlated well with each other; however, a significantly higher correlation (p<0.02) was demonstrated between the VerifyNow and Multiplate tests (RS=0.707; p<0.0001) than in other assay combinations (the VASP-assay and the VerifyNow device: RS=0.595; p<0.0001; the VASP-assay and the Multiplate analyzer: RS=0.588; p<0.0001). With respect to the recognition of high platelet reactivity, we found higher measurement concordance between the VerifyNow and Multiplate tests compared with other assay combinations, while for low platelet reactivity, only results of the VerifyNow and Multiplate assay were related to each other. Platelet reactivity measurements performed with the VASP, VerifyNow and Multiplate tests show comparably strong negative correlations with ticagrelor and AR-C124910XX concentrations.

Extrinsic blood coagulation pathway and risk factors for thrombotic events in patients with essential thrombocythemia.

INTRODUCTION    The clinical course of essential thrombocythemia (ET) is varied, and some patients do not exhibit any clinical signs of the disease at the time of diagnosis. The most frequent complications that occur during the course of ET are hemostasis abnormalities manifesting as hemorrhagic or thrombotic events. The mechanism of thrombotic events in patients with ET is complex and not fully understood. OBJECTIVES    The aim of the study was to evaluate the concentration and activity of tissue factor (TF) and tissue factor pathway inhibitor (TFPI), depending on the most important risk factors of thrombotic complications (age >60 years, history of thrombotic episodes, presence or absence of the JAK2 V617F mutation, and increased leukocyte count). PATIENTS AND METHODS    The study group included 113 patients with diagnosed ET, and the control group, 30 healthy volunteers matched for age and sex. The concentration and activity of TF and TFPI were measured using enzyme-linked immunosorbent assays. RESULTS    Patients with ET had a significantly higher activity and concentration of TF and increased activity of TFPI, as compared with controls. The analysis of the studied parameters in relation to risk factors revealed that patients with ET with a history of thrombotic events had a significantly higher concentration of TF, and patients with the JAK2 V617F mutation had a lower TFPI activity, as compared with patients without the mutation. CONCLUSIONS    Our study showed that in patients with ET who have a history of thrombosis or the JAK2 V617F mutation, the enhanced risk of thrombosis may result from an increased TF concentration or decreased TFPI activity.

Activation of the tissue factor-dependent extrinsic pathway and its relation to JAK2 V617F mutation status in patients with essential thrombocythemia.

Thrombotic complications may occur in 7.6-29.4% of patients with essential thrombocythemia. According to the cellular theory, tissue factor (TF) activating extrinsic blood coagulation pathway is essential for the activation of blood clotting. The aim of the study was to evaluate the activation of the TF-dependent extrinsic pathway in patients with essential thrombocythemia, depending on the presence or absence of the Janus kinase 2 (JAK2) V617F mutation. The study included 74 newly diagnosed patients (F/M: 47/27; mean age 61 years) with essential thrombocythemia (Tefferi and Vardiman, Leukemia 2008; 22(1):14-22). Patients were diagnosed in the Department of Clinical Hematology and Hematological Malignancies University Hospital No. 2, Bydgoszcz, Poland. The control group consisted of 30 healthy volunteers (F/M: 17/13; mean age 49 years). The concentration and activity of TF and TF pathway inhibitor (TFPI) were measured using ELISA method. In patients with essential thrombocythemia, we observed a higher concentration of TF [median (Me) = 686.90 vs 164.28 pg/ml] and over 10-fold higher activity of TF (Me = 46.05 vs 4.01 pmol/l) when compared with the control group. We also reported significantly higher activity of TFPI compared with the control group (Me = 1.93 vs 1.78 U/ml). Moreover, a concentration of TFPI was significantly lower in patients with essential thrombocythemia with JAK2 V617F mutation as compared with patients without the mutation (Me = 1.90 vs 2.16 U/ml; P = 0.039639). Increased TF activity and concentration is responsible for higher procoagulant potential in patients with essential thrombocythemia. Reduced activity of TFPI in patients with essential thrombocythemia with JAK2 V617F mutation indicates an increased prothrombotic risk in this group of patients.

CRP, but not TNF-α or IL-6, decreases after weight loss in patients with morbid obesity exposed to intensive weight reduction and balneological treatment.

OBJECTIVE: The aim of this study was to evaluate the concentrations of C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and the degree of homeostasis model assessment-insulin resistance (HOMA-IR) in patients with morbid obesity exposed to a three-week low-calorie diet and balneotherapy. METHODS: The study included 33 patients (25 females and 8 males; mean age 46 years) with body mass index (BMI) values of >40 kg/m(2). Evaluations of CRP, IL-6, TNF-α, lipid profile, HOMA-IR, and fasting glucose were carried out before (baseline data) and three weeks after the treatment. The control group consisted of 20 healthy volunteers (15 females and 5 males) with a mean age of 39 years and BMI values of ≤24.9 kg/m(2). RESULTS: In the blood of patients with morbid obesity we found significantly elevated levels of CRP, TNF-α, triglycerides, HOMA-IR and fasting glucose, but a decreased level of high density lipoprotein (HDL)-cholesterol, compared with the healthy individuals. The treatment resulted in about a 9.4% reduction in body weight from 122.5 to 111.0 kg and a significant decrease in the concentration of CRP, but no change in TNF-α or IL-6. HOMA-IR was significantly reduced. CONCLUSIONS: The decrease in CRP level without changes in TNF-α or IL-6 concentrations after the low-calorie diet and balneological treatment, suggests that an essential amount of adipose tissue must be removed before proper adipocyte function is restored. The decrease in HOMA-IR indicates an improvement in insulin sensitivity, which is beneficial in obese patients.

The number of circulating endothelial progenitor cells in healthy individuals--effect of some anthropometric and environmental factors (a pilot study).

PURPOSE: The aim of the study was the evaluation of the number of circulating endothelial progenitor cells (CEPCs) in healthy people and the assessment of the variability of quantitative of CEPCs after 6 weeks. MATERIAL AND METHODS: The study involved 48 healthy individuals; the group consisted of 24 men and 24 women; the mean age of 34. The criterion for the patients' eligibility for the study was the absence of diabetes, thrombosis and cardiovascular diseases such as atherosclerosis, hypertension, and heart failure. Neither did the respondents take any medication that could clearly affect the value of the results. In the whole blood samples the number of circulating endothelial progenitor cells was determined using flow cytometry. During the analysis the fluorescence of 100,000 cells was measured. CEPCs were identified with immunophenotype CD45-, CD31+, CD34+, CD133+. RESULTS: In the study, the median of the number of circulating endothelial progenitor cells in the whole group was 0.41/µL. There was also recorded an increased number of CEPCs after 6 weeks, as compared to the baseline; the difference was significant. There were no differences in the number of CEPCs between the women and the men. There was found no effect on the number of CEPCs factors such as: smoking, physical activity and alcohol consumption. CONCLUSIONS: The study showed that in healthy individuals the gender had no essential effect on the number of endothelial progenitor cells. Based on the demographic and lifestyle data acquired, it is difficult to explain the increase number of CEPCs after 6 weeks.

VEGF-A and PDGF-BB--angiogenic factors and the stage of diabetic foot syndrome advancement.

INTRODUCTION: In patients with diabetic foot syndrome (DFS), an inadequate angiogenic response is observed. The aim of this study was to evaluate the concentrations of VEGF-A, PDGF-BB, sVEGF-R2 and sVEGF-R1 in patients with diabetes-complicated diabetic foot syndrome and analyse them using selected clinical data. MATERIAL AND METHODS: Forty seven diabetic patients, 25 women mean age 63 and 20 men mean age 60.5, with diabetic foot syndrome (DFS) were enrolled in the experimental group. To evaluate angiogenesis factors depending on Wagner grade, the subjects were divided into three subgroups: I - patients with 0 Wagner grade (n = 14); II - patients with 1,2,3 Wagner grades (n = 15); and III - patients with 4,5 Wagner grades (n = 18). The control group consisted of 20 healthy volunteers. The material for research was blood. RESULTS: Significantly higher levels of VEGF-A and PDGF-BB in the DFS cases compared to controls were observed (VEGF-A p = 0.000001; PDGF-BB p = 0.000051). Analysis of angiogenic parameters according to the stage of diabetic foot syndrome advancement showed higher VEGF-A level (I: p = 0.000867; II: p = 0.001827; III: p = 0.000024) and PDGF-BB (respectively p = 0.004113, p = 0.004224, p = 0.002480) in all the subgroups. Decreased sVEGF-R2 concentrations were observed in the I (p = 0.054) subgroup and the III (p = 0.03524) subgroup. In this study, a strong positive correlation between VEGF-A and PDGF-BB was observed (R = 0.66; p = 0.000001). CONCLUSIONS: Our study revealed that proangiogenic factor levels were increased in DFS. This is associated with lower limb ischaemia and hypoxic conditions. The stage of diabetic foot syndrome advancement influenced VEGF-A and PDGF-BB concentrations.

Selected parameters of hemostasis in patients with myeloproliferative neoplasms.

Hemostatic disorders are a major clinical problem in patients with myeloproliferative neoplasms (MPNs) and they are the second most common cause of death in MPN patients, after infections. The aim of this study was to assess the fibrinolytic potential of the blood of patients with MPNs. The study involved 112 patients with MPNs diagnosed at the Hematology Clinic Dr J. Biziel University Hospital No. 2 in Bydgoszcz, Poland. The study group included 63 patients with essential thrombocythemia, 29 with polycythemia vera, 11 with chronic myelogenous leukemia (CML) and nine with primary myelofibrosis. The control group consisted of 25 healthy volunteers who were age and sex-matched. The following parameters were determined: concentration of tissue plasminogen activator antigen (t-PA:Ag), plasminogen activator inhibitor type 1 antigen concentration (PAI-1:Ag), D-dimer, thrombin-antithrombin complexes, fibrinogen, activated partial thromboplastin time and international normalized ratio. The study showed significantly increased t-PA:Ag, PAI-1:Ag and D-dimer levels in patients with MPNs. Moreover, we found increased concentrations of thrombin-antithrombin complexes and fibrinogen, as well as elevated platelet counts. Detailed analysis revealed that t-PA:Ag concentration was elevated in patients with essential thrombocythemia, CML and polycythemia vera. Concentration of PAI-1:Ag was increased in patients with essential thrombocythemia and polycythemia vera; D-dimer was significantly higher in essential thrombocythemia, polycythemia vera, CML and primary myelofibrosis patients. Increased concentrations of t-PA:Ag and D-dimer indicate secondary activation of the fibrinolytic system in patients with MPNs. Elevated levels of PAI-1 in MPN patients may result from its increased production by elevated number of activated platelets and vascular endothelial damage. PAI-1 by having an inhibitory effect on fibrinolysis manifests its procoagulant activity.

Comparison of serum prohepcidin and iron metabolism parameters in obese and non-obese elderly individuals.

INTRODUCTION: Current knowledge indicates that there is a close connection between being overweight, obesity and iron metabolism disorders,but the underlying mechanism is unclear. Hepcidin could be a major contributor to poor iron status observed in the obese population. MATERIAL AND METHODS: The study was performed in 58 obese elderly individuals (F/M 34/24) aged 65-91 (78.92 ± 8.32) years. The controlgroup consisted of 15 non-obese elderly volunteers, age- and sex-matched. Based on the WHO definition, 36 (62%) obese individualswere diagnosed with normo- or microcytic anaemia. The following parameters were determined: prohepcidin, haemoglobin, serum iron,erythropoietin, ferritin and C-reactive protein (CRP). RESULTS: Prohepcidin concentrations were significantly increased in obese elderly individuals without anaemia compared to obese andanaemic (p < 0.01) as well as non-obese volunteers (p < 0.01). In obese individuals with anaemia there was a decrease in serum iron,concomitant with increased levels of erythropoietin and CRP compared to two other groups. Ferritin concentration was increased inobese people (with and without anaemia) compared to the non-obese group. Serum prohepcidin levels were positively correlated withfat mass percentage in obese individuals without and with anaemia (r = 0.32; p = 0.02). CONCLUSION: Results of this preliminary study suggest that body fat content does have an impact on prohepcidin concentration, andthereby on iron homeostasis.

Serum prohepcidin and other iron metabolism parameters in elderly patients with anemia of chronic disease and with iron deficiency anemia.

INTRODUCTION: The diagnosis of iron deficiency anemia (IDA) in the context of the anemia of chronic disease (ACD) in elderly patients is often difficult due to the existence of many disorders. Recent studies have shown that hepcidin measurement (combined with the existing diagnostic methods) may possibly help in the differential diagnosis of IDA and ACD. OBJECTIVES: The aim of the study was to evaluate the differential diagnostic value of serum prohepcidin in elderly patients with IDA and ACD. PATIENTS AND METHODS: The study included 65 individuals aged 65 years or more: 26 patients with ACD, 13 patients with IDA, and 26 age-matched controls. Prohepcidin, ferritin, soluble transferrin receptor, erythropoietin, and interleukin 6 (IL-6) were measured using the commercially available enzyme-linked immunosorbent assay kits. Complete blood count, total iron-binding capacity (TIBC), and iron, transferrin, and C-reactive protein (CRP) levels were assayed using the standard laboratory methods. RESULTS: Prohepcidin concentrations were similar in patients with ACD (196.59 ng/ml) compared with those with IDA (230.16 ng/ml) (P = 0.35). Patients with ACD had significantly lower levels of TIBC compared with those with IDA (P <0.0001). Serum ferritin concentration in patients with ACD was almost 20-fold higher compared with those observed in patients with IDA (P <0.0001). CRP and IL-6 concentrations in patients with ACD were significantly higher compared with those with IDA. CONCLUSIONS: The results of the study indicate that serum prohepcidin has limited value in the differential diagnosis of IDA and ACD in elderly patients.

[Serum prohepcidin and other iron metabolism parameters in healthy adults]. / Ocena stezenia prohepcydyny na tle wybranych parametrów gospodarki zelazem w surowicy krwi ludzi zdrowych.

UNLABELLED: Hepcidin is a newly discovered iron metabolism regulator. It decreases iron absorption from the duodenal enterocytes and inhibits its release from the reticuloendothelial macrophages. The experiments on hepcidin and its precursor--prohepcidin seem very promising and underline the need to establish the reference intervals for these proteins and define the effects of age and sex on their concentrations. AIM: The aim of this study was to assess the impact of age and sex on serum prohepcidin concentration in healthy adults. MATERIAL AND METHODS: The study group consisted of 79 healthy volunteers (F-29, M-50) between 18 to 88 years of age. Healthy adults were divided into three groups according to age: group 1-33 healthy volunteers aged between 18-36, group 2-20 healthy volunteers aged between 38-53, group 3- 26 healthy volunteers aged between 58-88. Prohepcidin and ferritin concentrations were determined by enzyme-linked immunosorbent assays (ELISA). Serum iron concentration was measured on Architect c8000 System. RESULTS: In this work, we demonstrated that prohepcidin concentrations were significantly higher in group 3 (Me = 292,2 ng/ ml) in comparison with group 1 (Me = 150.7 ng/ml; p < 0.000001) or group 2 (Me = 118.1 ng/ml; p < 0.000001). We observed a positive correlation between age and prohepcidin concentration (R = 0.38; p = 0.001). The oldest group of volunteers also revealed significantly higher levels of ferritin and lower levels of serum iron in comparison with two groups of younger adults. Females showed significantly higher level of prohepcidin (Me = 226.51 ng/ml) than males (Me = 142.6 ng/ml; p = 0.01). We observed that females under 50 years of age had significantly higher levels of prohepcidin (Me = 290.9 ng/ml) in comparison with younger females (Me = 150.7 ng/ml; p = 0.0001), similar correlation was noticed in males (212.3 ng/ml vs 128.3 ng/ml; p = 0.007). CONCLUSIONS: Prohepcidin concentration increased significantly with age and was significantly higher in women than men. The results of our study show that serum prohepcidin concentration is age and sex dependent.