Publications by authors named "Ewa Żurawska-Płaksej"

13 Publications

  • Page 1 of 1

Platelet Reactivity and Response to Aspirin and Clopidogrel in Patients with Platelet Count Disorders.

Cardiol Res Pract 2021 17;2021:6637799. Epub 2021 Apr 17.

Department of Hematology, Blood Neoplasms, and Bone Marrow Transplantation, Wroclaw Medical University, Wrocław, Poland.

Background: Platelet reactivity and response to antiplatelet drugs, acetylsalicylic acid (ASA) and clopidogrel, in patients with thrombocytopenia and thrombocythemia can have a potentially important effect on the outcome. The effectiveness and safety of antiplatelet drugs in such patients has not been well examined. Measuring the effect of ASA and clopidogrel on platelets could help guide the therapy. Nevertheless, platelet response to antiplatelet drugs is not routinely measured in platelet count disorders and relevant evidence is scarce.

Aims: The study aimed to measure platelet reactivity and response to ASA and clopidogrel in patients with platelet count disorders.

Materials And Methods: This was a cross-sectional study of consecutive patients hospitalized in cardiology and hematology departments in the years 2018-2019. The study included patients with thrombocytopenia (PLT < 150 G/L) and thrombocythemia (PLT > 450 G/L) on ASA or dual antiplatelet therapy (DAPT; ASA plus clopidogrel). Controls included patients on antiplatelet drugs with normal platelet count. Platelet reactivity was measured in whole blood (Multiplate aggregometer, Roche, Switzerland) using arachidonic acid (AA), adenosine-5'-diphosphate (ADP), and thrombin receptor agonist peptide-6 (TRAP) as agonists. Platelet aggregation was expressed in arbitrary units (AU). AA-induced aggregation was used as a measure of response to ASA with a cut-off above 30 AU showing high on-treatment platelet reactivity to ASA (HTPR-A). ADP-induced aggregation measured response to clopidogrel with a cut-off above 48 AU for high on-treatment platelet reactivity to clopidogrel (HTPR-C). TRAP-induced aggregation measured baseline platelet reactivity not affected by oral antiplatelet drugs.

Results: The study included 174 patients. There were 64 patients with thrombocytopenia, 30 patients with chronic thrombocythemia, and 80 controls. All patients were on 75 mg of ASA and 32% of them additionally on 75 mg of clopidogrel due to a history of recent coronary artery angioplasty. AA- and ADP-induced aggregation was comparable between thrombocytopenic patients and controls (median (IQR) 19 (7-28) vs. 23 (15-38) for AA AU and 32 (16-44) vs. 50 (32-71) for ADP AU, respectively), while it was significantly higher in thrombocythemic patients (median (IQR) 80 (79-118) for AA AU and 124 (89-139) for ADP AU). TRAP-induced aggregation showed significantly lowest aggregation in thrombocytopenic (median (IQR) 41 (34-60) for TRAP AU) and highest in thrombocythemic patients (median (IQR) 137 (120-180) for TRAP AU). HTPR-A was frequent in thrombocythemic patients in comparison with thrombocytopenic patients and controls (60% vs. 4% vs. 15%, respectively; < 0.0002). HTPR-C was highly common in thrombocythemic patients and least common in thrombocytopenic ones in comparison with controls (80% vs. 8% vs. 40%, respectively; < 0.001).

Conclusion: Chronic thrombocytopenia does not significantly affect platelet reactivity and response to ASA and clopidogrel in comparison with controls. Thrombocytosis significantly increases platelet reactivity and attenuates response to both ASA and clopidogrel.
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http://dx.doi.org/10.1155/2021/6637799DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8068533PMC
April 2021

Platelet polyphosphate level is elevated in patients with chronic primary thrombocytopenia: A preliminary study.

Adv Clin Exp Med 2020 Sep;29(9):1051-1056

Department of Pharmaceutical Biochemistry, Wroclaw Medical University, Poland.

Background: Platelets are key players in hemostasis. These blood cells contain different types of granules. Recently, there has been a growing interest in the role of inorganic polyphosphate (polyP) structures stored in dense granules of platelets and secreted during platelet activation.

Objectives: To measure platelet polyP levels in patients with thrombocytopenia and thrombocythemia, and to examine the relationship of this indicator with platelet aggregation.

Material And Methods: The study included 36 patients with hematological disorders (26 with primary chronic thrombocytopenia and 10 with essential thrombocythemia (ET)) and 40 healthy subjects. Platelet reactivity was measured using whole blood impedance aggregometry. The polyP levels were isolated from lysed platelets, which were obtained from citrated platelet-rich plasma. The procedure included inactivating endogenous phosphatases, removing phosphate units derived from DNA and proteins, and finally hydrolyzing them into monophosphate units. A colorimetric assay using malachite green and ammonium molybdate was performed in order to quantify polyP levels.

Results: The polyP concentrations were significantly higher in the patients with thrombocytopenia than in the patients with thrombocythemia or the controls. The polyP level was not correlated with the level of aggregation.

Conclusions: The higher polyP levels observed in the patients with low platelet counts may indicate the existence of a compensatory mechanism that prevents excessive bleeding in such patients. Our study provides evidence of an essential role of polyP in platelet function and the coagulation process.
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http://dx.doi.org/10.17219/acem/125430DOI Listing
September 2020

Parameters of Oxidative and Inflammatory Status in a Three-Month Observation of Patients with Acute Myocardial Infarction Undergoing Coronary Angioplasty-A Preliminary Study.

Medicina (Kaunas) 2019 Sep 13;55(9). Epub 2019 Sep 13.

Department of Toxicology, Wroclaw Medical University, 50-556 Wroclaw, Poland.

: Patients with acute myocardial infarction (MI) are usually treated with percutaneous transluminal coronary angioplasty (PTCA), which is burdened with a risk of postoperative complications, often accompanied by biochemical disturbances. The aim of our study was to evaluate a set of selected parameters of oxidative and inflammatory status, which could be useful in the management of post-procedural care in MI patients after PTCA. : In this preliminary study, ischemia modified albumin (IMA), advanced oxidation protein products (AOPP), thiol groups (SH), total antioxidant status (TAS), insulin growth factor-1 (IGF-1), presepsin (PSP), and trimethylamine N-oxide (TMAO) were chosen as candidate biomarkers, and were determined in patients with MI who underwent PTCA at two time points: During cardiac episodes (at admission to the hospital, T0) and 3 months later (T3). : Most of the examined parameters were significantly different between patients and control subjects (except for IMA and TAS), but only hsCRP changed significantly during the time of observation (T0 vs. T3). Discriminant analysis created a model composed of AOPP, hsCRP, PSP, and TMAO, which differentiated male subjects into a group with MI and a control (without cardiovascular diseases). : This set of parameters seems useful in evaluating inflammatory and oxidative status in MI patients after PTCA.
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http://dx.doi.org/10.3390/medicina55090585DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6780791PMC
September 2019

The effect of glycation on bovine serum albumin conformation and ligand binding properties with regard to gliclazide.

Spectrochim Acta A Mol Biomol Spectrosc 2018 Jan 6;189:625-633. Epub 2017 Sep 6.

Department of Toxicology, Faculty of Pharmacy, Wroclaw Medical University, Borowska 211, 50-556 Wroclaw, Poland. Electronic address:

Albumin, the major serum protein, plays a variety of functions, including binding and transporting endogenous and exogenous ligands. Its molecular structure is sensitive to different environmental modifiers, among which glucose is one of the most significant. In vivo albumin glycation occurs under physiological conditions, but it is increased in diabetes. Since bovine serum albumin (BSA) may serve as a model protein in in vitro experiments, we aimed to investigate the impact of glucose-mediated BSA glycation on the binding capacity towards gliclazide, as well as the ability of this drug to prevent glycation of the BSA molecule. To reflect normo- and hyperglycemia, the conditions of the glycation process were established. Structural changes of albumin after interaction with gliclazide (0-14μM) were determined using fluorescence quenching and circular dichroism spectroscopy. Moreover, thermodynamic parameters as well as energy transfer parameters were determined. Calculated Stern-Volmer quenching constants, as well as binding constants for the BSA-gliclazide complex, were lower for the glycated form of albumin than for the unmodified protein. The largest, over 2-fold, decrease in values of binding parameters was observed for the sample with 30mM of glucose, reflecting the poorly controlled diabetic state, which indicates that the degree of glycation had a critical influence on binding with gliclazide. In contrast to significant changes in the tertiary structure of BSA upon binding with gliclazide, only slight changes in the secondary structure were observed, which was reflected by about a 3% decrease of the α-helix content of glycated BSA (regardless of glucose concentration) in comparison to unmodified BSA. The presence of gliclazide during glycation did not affect its progress. The results of this study indicate that glycation significantly changed the binding ability of BSA towards gliclazide and the scale of these changes depended on glucose concentration. It may have a direct impact on the free drug fraction and its pharmacokinetic behavior, including the risk of hypoglycemic episodes or unexpected interactions with other ligands. The use of BSA in examining binding effects upon glycation seems to be good model for preliminary research and may be used to identify a potential drug response in a diabetic state.
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http://dx.doi.org/10.1016/j.saa.2017.08.071DOI Listing
January 2018

Increased chitotriosidase activity in plasma of patients with type 2 diabetes.

Arch Med Sci 2016 Oct 20;12(5):977-984. Epub 2016 May 20.

Department of Toxicology, Wroclaw Medical University, Wroclaw, Poland.

Introduction: Chitotriosidase (CHIT1) is a chitinolytic enzyme involved mainly in the immune and inflammatory response. It shows increased activity in many pathologies, including in newly diagnosed type 2 diabetes (T2D). This study aimed to investigate this enzyme's activity in plasma of patients with ongoing T2D and indicate factors related to the increased activity of this enzyme.

Material And Methods: Ninety-one patients and 46 control subjects without abnormalities in carbohydrate metabolism and inflammatory states were enrolled in the study. Plasma CHIT1 activity was measured by a spectrofluorometric method. Routine laboratory parameters such as blood glucose, total cholesterol and HDL fraction, triglyceride, glycated hemoglobin, white blood cell count and C-reactive protein were measured by standard methods.

Results: We found that the chitotriosidase activity was significantly higher ( < 0.001) in type 2 diabetic patients and positively associated with parameters of glycemic control (levels of glucose and glycated hemoglobin) and blood pressure. Plasma glucose level and systolic blood pressure were independent determinants of increased CHIT1 activity in T2D patients, even after adjustment for disease duration, body mass index, parameters of inflammation and lipid metabolism. We also found that increased CHIT1 activity was associated with occurrence of diabetic angiopathies.

Conclusions: This investigation indicates a possible role of chitotriosidase in the course of T2D, especially in relation to development of diabetic angiopathies.
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http://dx.doi.org/10.5114/aoms.2016.60093DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5016580PMC
October 2016

Neutrophils as a Source of Chitinases and Chitinase-Like Proteins in Type 2 Diabetes.

PLoS One 2015 30;10(10):e0141730. Epub 2015 Oct 30.

Department of Toxicology, Wroclaw Medical University, Wroclaw, Poland.

Purpose: The pathophysiological role of human chitinases and chitinase-like proteins (CLPs) is not fully understood. We aimed to determine the levels of neutrophil-derived chitotriosidase (CHIT1), acidic mammalian chitinase (AMCase) and chitinase 3-like protein 1 (YKL-40) in patients with type 2 diabetes (T2D) and verify their association with metabolic and clinical conditions of these patients.

Methods: Neutrophils were obtained from the whole blood by gradient density centrifugation from 94 T2D patients and 40 control subjects. The activities of CHIT1 and AMCase as well as leukocyte elastase (LE) were measured fluorometrically and concentration of YKL-40 immunoenzymatically. Also, routine laboratory parameters in serum/plasma were determined by standard methods.

Results: The levels of all three examined proteins were about 2-times higher in diabetic patients in comparison to control subjects. They were significantly correlated with the activity of LE and increased progressively across tertiles of LE activity. Moreover, the activities of CHIT1 and AMCase were significantly correlated with each other. Metabolic compensation of diabetes did not influence the levels of these proteins. In the subgroup of patients with inflammatory evidence only YKL-40 concentration was significantly higher compared to those without inflammation. The highest levels of all three proteins were observed in patients with macroangiopathies. Insulin therapy was associated with lower levels of examined proteins.

Conclusions: We revealed that neutrophils may be an important source of the increased levels of chitinases and CLPs in T2D, and these proteins may participate in inflammatory mechanisms in the course of the disease and consequent development of diabetic angiopathies.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0141730PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4627817PMC
June 2016

Changes in glycosylation of human blood plasma chitotriosidase in patients with type 2 diabetes.

Glycoconj J 2016 Feb;33(1):29-39

Human blood plasma chitotriosidase (CHIT1) is a glycoprotein with chitinolytic activity with not fully elucidated biological function. Its increased level is observed in type 2 diabetes mellitus (T2DM) and is associated with development of diabetic complications. The CHIT1 glycosylation profile and degree is still poorly studied and never investigated in T2DM. Therefore the aim of the present study was to examine the association between glycosylation profile and degree and diabetes with accompanying nephropathy. In blood plasma of 28 patients with T2DM and 11 healthy subjects the CHIT1 concentration and specific activity were examined. The profile and degree of CHIT1 glycosylation were determined by lectin-ELISA using lectins specific to O-glycans (Jacalin, MPL, VVL) and sialo-specific SNA and MAA. We revealed that both concentration and specific activity of CHIT1 significantly increased in T2DM, especially in nephropathy with elevated albuminuria. The relative reactivities with lectins, except Jacalin, decreased progressively with T2DM occurrence and albuminuria progression. The most significant differences were observed between control vs. albuminuric group (Micro and Macro). It is also possible that the observed differences in immunoblotting pattern in molecular masses of CHIT1 bands between T2DM patients and healthy subjects may be caused by the differences in degree of CHIT1 glycosylation. The analysis of CHIT1 glycosylation status and the determination of CHIT1 concentration together with its enzymatic activity in blood plasma might constitute additional valuable diagnosis tools for the evaluation the T2DM patients with accompanying nephropathy. Extension of the lectin panel specific to O-glycans occurs useful for the further research using microarray formats, which are expected to accelerate “lectin-based glycan profiling” of glycoproteins.
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http://dx.doi.org/10.1007/s10719-015-9629-zDOI Listing
February 2016

[Glycated albumin as a marker of glycemia in diabetes and its vascular complications].

Postepy Hig Med Dosw (Online) 2015 May 17;69:638-48. Epub 2015 May 17.

Katedra i Zakład Biochemii Farmaceutycznej, Wydział Farmaceutyczny z Oddziałem Analityki Medycznej, Uniwersytet Medyczny im. Piastów Śląskich we Wrocławiu.

Effective glycemic control is very important to prevent the onset and the progression of chronic complications in diabetic patients. It is known that glycation of various proteins is increased in diabetic patients compared with non-diabetics. Among these glycated proteins, glycated hemoglobin (HbA1c) is commonly used as a gold standard index of glycemic control in the clinical setting. However, it can be unreliable in conditions affecting the lifespan of erythrocytes (120 days) as well as in the clinical state in which glycemic control alleviates or deteriorates in a short period. By overcoming the shortcomings of HbA1c, glycated albumin (GA) has gained interest as a useful index for an intermediate glycation period (2 weeks) and pathogenic protein. After giving a brief overview of the key role of HbA1c as a long-term glycemic marker, this review focuses on (a) glycation of human albumin and its main properties, (b) methods of GA determination, (c) the recent clinical status of GA as a glycemic index in diabetic patients and its association with vascular complications. Finally, conditions with a possible inaccurate GA level are also mentioned.
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http://dx.doi.org/10.5604/17322693.1153082DOI Listing
May 2015

Proteins from the 18 glycosyl hydrolase family are associated with kidney dysfunction in patients with diabetes type 2.

Biomarkers 2015 Feb 18;20(1):52-7. Epub 2014 Dec 18.

Department of Pharmaceutical Biochemistry, Wroclaw Medical University , Wroclaw , Poland .

Objectives: To investigate chitotriosidase (CHIT1) activity and chitinase-3-like protein 1 (YKL-40) concentration in plasma of type 2 diabetic patients and evaluate their relationship with kidney dysfunction.

Materials And Methods: 94 diabetic subjects and 33 controls were enrolled in the study. Plasma CHIT1 activity and YKL-40 concentration were measured along with routine laboratory parameters.

Results: Levels of CHIT1 and YKL-40 in plasma of type 2 diabetic patients increased progressively with the degree of albuminuria. CHIT1 discriminated normoalbuminuric subjects from those with abnormal albuminuria better than YKL-40.

Conclusions: CHIT1represent a supportive biomarker connected with development of diabetic vascular complications, especially kidney dysfunction.
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http://dx.doi.org/10.3109/1354750X.2014.992475DOI Listing
February 2015

Activities of neutrophil membrane-bound proteases in type 2 diabetic patients.

Arch Med Res 2014 Jan 5;45(1):36-43. Epub 2013 Dec 5.

Department of Pharmaceutical Biochemistry, Wroclaw Medical University, Wroclaw, Poland.

Background And Aims: Hyperglycemia and oxidative stress in type 2 diabetes (T2DM) provoke neutrophil overstimulation and the release and/or translocation of proteases from granules to the cell surface. Although the expression of neutrophil membrane-bound elastase (MLE) is well documented, the presence of the membrane-bound form of cathepsin B (MCB) is unknown. The aim of our study was to evaluate the neutrophil MLE and MCB activities in T2DM patients and their associations with the metabolic and clinical parameters of the disease.

Methods: Neutrophils were obtained from 47 T2DM patients and 20 control subjects. The activities of MLE and MCB and the intracellular activities of the examined proteases (ILE and ICB, respectively) were measured using fluorometric substrates. Additionally, the percentage equivalents of the activities, namely, MLEtot/ILEtot and MCBtot/ICBtot, were calculated. The susceptibility to inhibitors of both forms of the studied proteases was also determined.

Results: A significant increase in the activities of MLE, MCB, ILE, and ICB was found in neutrophils from T2DM patients compared with the control group. The percentage equivalent (contribution of the total membrane-bound activities to the total intracellular activities) was also higher. A partial resistance of the membrane-bound forms toward their inhibitors was revealed. Higher activities of both the membrane-bound and the intracellular proteases were also observed in patients with poor glycemic and metabolic control. The differences between subgroups with different therapeutic schemes were also revealed.

Conclusions: The pathophysiological implications of the neutrophil membrane-bound forms of leukocyte elastase and cathepsin B are of great importance in the development of T2DM and its complications.
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http://dx.doi.org/10.1016/j.arcmed.2013.10.003DOI Listing
January 2014

Solid dispersion in pharmaceutical technology. Part II. The methods of analysis of solid dispersions and examples of their application.

Polim Med 2012 ;42(2):97-107

Department of Drug Form Technology, Wroclaw Medical University, Wroclaw, Poland.

In the first part of the article solid dispersions were classified the properties and methods of their preparation were described. This section presents methods of analysis of solid dispersions i.e.: thermoanalytical methods, XRPD, FTIR, microscopic methods, dissolution studies and examples of drug forms where solid dispersions were used.
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October 2012

Solid dispersions in pharmaceutical technology. Part I. Classification and methods to obtain solid dispersions.

Polim Med 2012 ;42(1):17-27

Department of Drug Form Technology, Wroclaw Medical University, ul. Szewska 38, Wroclaw, Poland.

There are many methods to increase solubility of a substance. These include, inter alia, preparation of solid dispersions, i.e. eutectic mixtures, solid solutions, glassy solutions and suspensions. When compared to the individual constituents prior to dispersion formation solid dispersion components are better soluble in water. Therefore, solid solutions became one of the most promising ways to modify solubility, ensuring improved bioavailability and consequently therapeutic efficacy of a substance. In this part of the publication solid dispersions were classified and described in regard to their properties and preparation methods, i.e. melting method, melt evaporation and melt extrusion methods, lyophilisation technique, melt agglomeration process as well as SCF technology and electrospinning.
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August 2012

[Ischemia modified albumin--specific marker in cardiological diagnostics?].

Wiad Lek 2008 ;61(10-12):263-8

Katedry i Kliniki Angiologii, Nadciśnienia Tetniczego i Diabetologii oraz, Akademii Medycznej we Wrocławiu.

Ischemia modified albumin (IMA) is a new biological marker for early identification of chest pain and ruling out myocardial infarction among patients with acute syndromes submitting to emergency department. Recently IMA has been investigated in the light of other cardiac markers (cTnT, CK-MBmas, NT-proBNP) in various states of ischemia (acute coronary syndromes, after percutaneous coronary intervention, in coronary vasospasm). Ischemia modified albumin levels were elevated in these states what suggests myocardial ischemia. However decrease in IMA concentration after exercise-induced skeletal muscle ischemia still remains unclear. Increased IMA concentration in patients with acute ischemic stroke and exposed to trauma limits its ability for detection myocardial ischemia. Specificity of IMA measurement is limited also in patients with peripheral vascular disease, systemic sclerosis, diabetes, end stage renal disease, pulmonary embolism and other pathological states with accompanying oxidative stress.
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April 2009