Regional Center for Transfusion Medicine

Białystok, Poland

Regional Center for Transfusion Medicine

Białystok, Poland
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Ratajczak-Wrona W.,Medical University of Bialystok | Jablonska E.,Medical University of Bialystok | Garley M.,Medical University of Bialystok | Jablonski J.,Medical University of Bialystok | And 2 more authors.
Journal of Immunotoxicology | Year: 2013

The aim of the study was to assess the activity of AP-1 family proteins, e.g. Fra-1, Fra-2, JunB, JunD, and FosB, engaged in the regulation of inducible nitric oxide synthase (iNOS) expression and the production of NO by neutrophils (PMN) exposed to N-nitrosodimethylamine (NDMA) xenobiotic. Isolated human PMN were incubated in the presence of NDMA. iNOS mRNA expression was then analyzed using Northern blot and the expression of other proteins in the cytoplasmic and nuclear fractions were assessed using Western blot. The obtained results indicate that NDMA increased iNOS mRNA and protein expression in human PMN. Furthermore, it increased the expression of Fra-1, Fra-2, JunB, and JunD in the cytoplasmic fraction, and FosB expression in the fractions of analyzed cells. As a consequence of inhibiting p38 pathway and JNK, reduced iNOS expression and NO production was noted in PMN exposed to NDMA. Inhibition of the p38 pathway resulted in reduced expression of all analyzed proteins in the cytoplasmic fraction of PMN exposed to NDMA. Furthermore, increased Fra-2 expression and reduced FosB expression were found in the nuclear fraction of those cells. Inhibiting ERK5 pathway resulted in increased JunB expression in both fractions of the analyzed cells. Therefore, no changes in the expression of analyzed proteins in the presence of NDMA were observed in PMN pre-incubated with JNK pathway inhibitor. In conclusion, the results here indicate a role of Fra-1, Fra-2, JunB, JunD, and FosB transcription factors in the regulation of iNOS expression and NO production by human neutrophils exposed to NDMA. © 2013 Informa Healthcare USA, Inc.


Ciborowski M.,University of San Pablo - CEU | Ciborowski M.,Medical University of Bialystok | Javier Ruperez F.,University of San Pablo - CEU | Martinez-Alcazar M.P.,University of San Pablo - CEU | And 6 more authors.
Journal of Proteome Research | Year: 2010

Professional and recreational diving are growing activities in modern life. Diving has been associated with increased prevalence of stroke, hypertension, asthma, diabetes, or bone necrosis. We evaluated the effect of increased pressure equivalent to diving at 30 and 60 m for 30 min in two groups of divers using an untargeted approach with LC-MS fingerprinting of plasma. We found over 100 metabolites to be altered in plasma post exposure and after the corresponding decompression procedures. Among them, a group of lysophosphatidylcholines and lysophosphatidylethanolamines were increased, including lysoplasmalogen, a thrombosis promoter, together with changes in metabolic rate-associated molecules such as acylcarnitines and hemolysis-related compounds. Moreover, three metabolites that could be associated to bone degradation show different intensities between experimental groups. Ultimately, this nontargeted, short-term study opens the possibility of discovering markers of long-term effect of pressure that could be employed in routine health control of divers and could facilitate the development of safer decompression procedures. © 2010 American Chemical Society.


PubMed | University of Warmia and Mazury, National Health Research Institute, Medical University of Bialystok, Medical University of Lublin and Regional Center for Transfusion Medicine
Type: | Journal: BMC cancer | Year: 2015

Of many specialized blood cells, monocytes are gaining increasing attention for their role in neoplastic disorders. The purpose of the present investigation was to determine the expression of selected peripheral blood monocyte surface antigens in cases of cervical, endometrial, and ovarian cancers. In addition, our aim was to validate the diagnostic value of two artificial coefficients recently proposed for the diagnosis of gynecologic malignancies: Neutrophil to Lymphocyte Ratio (NLR), and Multiplication of Neutrophil and Monocyte Counts (MNM).We studied 69 white Caucasian women with histopathologic confirmation of endometrial (N=42), cervical (N=13), and ovarian (N=14) cancers. Reference Group I were women suspected of cancer but histologically nullified (N=20), and Group II were healthy blood donors (N=23). Expression of CD11a, CD11b, CD11c, CD16, CD54 (ICAM-1), CD62 L (L-selectin), CD64, and HLA-DR was measured with immunofluorescence in a flow cytometer.CD54 expression increased by 35.6% (p<0.001) whilst HLA-DR decreased by 10.8% (p<0.001) in all cancer subgroups and Group I as compared to blood donors. A correlation (p<0.05) between CD54 and CD62 L was stronger in all cancers studied than in healthy subjects. There was no difference in the NLR values between any of these subgroups. Moreover, we observed an increase in MNM parameter in cases of cervical and endometrial cancer and in the Reference Group I.In the studied gynecologic malignancies, CD54 expression on peripheral blood monocytes is enhanced, indicating a higher transmigrational potential present in such patients, and HLA-DR expression diminished, indicating a decreased readiness of the immune system to recognize foreign antigens. The more pronounced correlation for the expression of CD54 and CD62 L in cancer suggests that monocytes uptake from the bloodstream and their local adhesion increase the pool of tumor-associated macrophages. This study challenged the suggested credibility and usefulness of the artificial parameters of MNM and NLR for the differential diagnosis of gynecologic malignancies.


Ratajczak-Wrona W.,Medical University of Bialystok | Jablonska E.,Medical University of Bialystok | Garley M.,Medical University of Bialystok | Jablonski J.,Medical University of Bialystok | Radziwon P.,Regional Center for Transfusion Medicine
Indian Journal of Experimental Biology | Year: 2013

Potential role of ERK1/2 kinase in conjunction with p38 in the regulation of inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production, and superoxide anion generation by human neutrophils (PMNs) exposed to N-nitrosodimethylamine (NDMA) was determined. Increased synthesis of NO due to the involvement of iNOS in neutrophils exposed to NDMA was observed. In addition, intensified activation of ERK1/2 and p38 kinases was determined in these cells. Inhibition of kinase regulated by extracellular signals (ERK1/2) pathway, in contrast to p38 pathway, led to an increased production of NO and expression of iNOS in PMNs. Moreover, as a result of inhibition of ERK1/2 pathway, a decreased activation of p38 kinase was observed in neutrophils, while inhibition of p38 kinase did not affect activation of ERK1/2 pathway in these cells. An increased ability to release superoxide anion by the studied PMNs was observed, which decreased after ERK1/2 pathway inhibition. In conclusion, in human neutrophils, ERK1/2 kinase is not directly involved in the regulation of iNOS and NO production induced by NDMA; however, the kinase participates in superoxide anion production in these cells.


Ratajczak-Wrona W.,Medical University of Bialystok | Jablonska E.,Medical University of Bialystok | Garley M.,Medical University of Bialystok | Jablonski J.,Medical University of Bialystok | And 3 more authors.
Advances in Medical Sciences | Year: 2013

Purpose: The role of MAP kinases in the activation of AP-1 (c-Jun, c-Fos) and NF-κB p65 engaged in the regulation of iNOS expression in human neutrophils (PMNs) exposed to N-nitrosodimethylamine (NDMA) was analyzed in the study. Material and Methods: The study included a group of 20 healthy individuals. Isolated human PMN were incubated in the presence of NDMA. Selective MAP kinases inhibitors were used. The expression of proteins in the cytoplasmic and nuclear fractions was assessed using Western blot method. Results: The results show that NDMA intensifies iNOS, c-Jun, NF-κB p65 and IκB-α expression in the analyzed PMNs. The blocking of the p38 pathway led to lower iNOS expression, and higher expression of c-Jun and c-Fos in the cytoplasmic fraction, and also lower c-Jun expression in the nuclear fraction of PMNs exposed to NDMA. A decrease in iNOS expression in the cytoplasmic fraction, and also c-Jun in both fractions of the examined cells, was observed as a result of JNK pathway inhibition. The blocking of the ERK5 pathway led to higher iNOS, c-Jun and c-Fos expression in the cytoplasmic fraction, and higher c-Jun expression in the nuclear fraction of PMNs exposed to NDMA. The study also demonstrated that blocking of the p38 and JNK pathways resulted in higher expression of NF-κB p65 and IκB-α in the cytoplasmic fraction and their lower expression in the nuclear fraction of these cells. Conclusion: Our data indicate the role of MAP kinases p38 and JNK in the activation of c-Jun and NF-κB p65 transcription factors engaged in the regulation of iNOS expression in human neutrophils exposed to NDMA. However ERK5 kinase is not involved in the regulation of iNOS and NO production by those cells. © Medical University of Bialystok, Poland.


Ratajczak-Wrona W.,Medical University of Bialystok | Jablonska E.,Medical University of Bialystok | Garley M.,Medical University of Bialystok | Jablonski J.,Medical University of Bialystok | Radziwon P.,Regional Center for Transfusion Medicine
APMIS | Year: 2011

In neutrophils (PMN) and mononuclear cells (PBMC), one of the enzymes responsible for nitric oxide (NO) synthesis is inducible nitric oxide synthase (iNOS). Changes in iNOS expression result from various signalling pathways including the mitogen-activated protein kinase (MAPK) pathway activated by endogenic and exogenic factors. N-nitrosodimethylamine (NDMA) is a xenobiotic with widespread occurrence in human environment and has an effect on cells of the immune system. The aim of this study was to determine the effect of NDMA on iNOS expression and NO production by human PMN and PBMC cells in the light of superoxide anion production by PMN cells. Moreover, the role of JNK and p38 pathways in NO production with involvement of iNOS was studied. Additionally, the function of JNK pathway in generation of superoxide anion was determined. Moreover, nitrotyrosine expression was studied in PMN and PBMC cells in the presence of NDMA. This work shows that NDMA increases iNOS expression and NO production by PMN and PBMC cells. In addition, elevated expression of phospho-JNK and phospho-p38, which are markers of JNK and p38 MAPK pathways activation, were observed. Lower iNOS expression and NO production by neutrophils exposed to extended action of NDMA were observed after application of inhibitor of JNK and p38 pathways. Lower phospho-p38 expression in PMN stimulated by NDMA was found as a result of arresting JNK pathway, whereas, application of inhibitor of p38 pathway resulted in enhanced phospho-JNK expression in PMN and PBMC cells. Increased ability to release superoxide anion by NDMA-stimulated PMN cells was observed. This ability was reduced after the application of inhibitor of JNK pathway. In PMN and PBMC cells exposed to NDMA, an increased expression of nitrotyrosine, which is dependant on JNK and p38 pathways that are activated by this particular xenobiotic, was observed. Generally, increased induction of iNOS related to elevated production of NO by PMN and PBMC cells exposed to NDMA may result in dysfunction of regulation of immunity responses controlled by this molecule in various conditions. Increased expression of nitrotyrosine in PMN and PBMC cells exposed to NDMA may affect their functions in an auto- and/or a paracrine way. Mutual interactions of JNK and p38 MAPK during the induction of iNOS expression in cells exposed to NDMA indicate complex mechanism of induction of iNOS synthase. © 2011 The Authors. APMIS © 2011 APMIS.


Rusak T.,Medical University of Bialystok | Ciborowski M.,Medical University of Bialystok | Uchimiak-Owieczko A.,Regional Center for Transfusion Medicine | Piszcz J.,Medical University of Bialystok | And 3 more authors.
Platelets | Year: 2012

Polycythemia vera (PV) is associated with an increased frequency of thrombotic complications. This study was undertaken to evaluate the hemostatic balance in the blood of PV patients by means of thromboelastography (TEG). The effect of isovolemic erythrocytapheresis (ECP) on the hemostasis of PV patients was also studied. We assessed the coagulation status of 76 PV patients undergoing ECP and 50 of healthy controls. TEG measurements were performed immediately before and after the ECP procedure. Coagulation was triggered by recalcification in freshly collected citrated blood. We recorded clotting time (R), alpha angle, and maximum amplitude (MA) of the clot. The results presented here show that, compared with healthy controls, PV patients demonstrated an increase in alpha angle (p<0.005) and in MA (p=0.14). In the subgroup of PV patients with high (>440×109l-1) platelet (PLT) count, differences in MA (p<0.01) and alpha angle (p<0.001) were more significant. Following ECP procedure, a significant (p≤0.01) reduction of R time, a rise of alpha angle, and MA were observed, indicating augmentation of a hypercoagulable state. In PV patients, the rise in alpha angle positively correlated (r=0.549) with platelet count but not with the number of erythrocytes and leukocytes. Following ECP, this correlation was reduced (r=0.382). Dilution (with saline) of blood from PV patients and of healthy controls, to a degree similar to that used during the ECP procedure, resulted in reduction of R and rise of the alpha angle. In conclusion, TEG measurements show that the majority of PV patients demonstrate abnormal hemostasis in which a major role is played by platelets rather than plasma factors. The hypercoagulable state in PV patients is significantly augmented following the ECP and may be related to the hemodilution intrinsically included in this procedure. TEG may help to assess the thrombotic risk in individual PV patients. © 2012 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.


Ratajczak-Wrona W.,Medical University of Bialystok | Jablonska E.,Medical University of Bialystok | Garley M.,Medical University of Bialystok | Jablonski J.,Medical University of Bialystok | And 2 more authors.
Turkish Journal of Biology | Year: 2013

The study objective was to assess the role of the JAK/STAT pathway in the induction of nitric oxide (NO) synthesis with the involvement of inducible nitric oxide synthase (iNOS) and in the generation of superoxide anion radical by human neutrophils (PMNs) exposed to N-nitrosodimethylamine (NDMA). Isolated PMNs were incubated in the presence of NDMA. The expression of the study proteins in the cytoplasmic and nuclear fraction was determined by western blotting. Total NO was assayed in cell cultures by Griess reaction. The ability of PMNs to generate superoxide anion radical was assessed using the cytochrome-c reduction test. The current results indicate that NDMA potentiates the expression of iNOS and phospho-JAK2 in the cytoplasmic fraction of human PMNs. Moreover, it increases the expression of phospho-STAT1 and phospho-STAT3 in both fractions of the cells studied. PMNs treated with NDMA showed greater capability to release NO and superoxide anion radical, which was reduced after administration of the JAK/ STAT pathway inhibitor. The current findings indicate that NDMA in PMNs activates the JAK/STAT pathway, which plays a role in the production of NO with the involvement of iNOS and in the generation of superoxide anion radical by these cells. © TÜBITAK.


Ratajczak-Wrona W.,Medical University of Bialystok | Jablonska E.,Medical University of Bialystok | Garley M.,Medical University of Bialystok | Jablonski J.,Medical University of Bialystok | And 3 more authors.
Journal of Immunotoxicology | Year: 2014

Neutrophils (PMN) play diverse regulatory and effector functions in the immune system through the release of reactive nitrogen species, including nitric oxide (NO). The enzyme responsible for NO synthesis in PMN is inducible nitric oxide synthase (iNOS) that is regulated by various signaling pathways, e.g. PI3K-Akt/PKB, and transcription factors. N-Nitrosodimethylamine (NDMA), a xenobiotic widespread in the human environment, affects immune cells. The study objective here was to examine the role of the PI3K-Akt/PKB pathway in induction of NO synthesis (with involvement of iNOS) in human PMN, as well as in autologous mononuclear cells (PBMC), exposed to NDMA. Isolated cells were incubated for 2h with a sub-lethal dose of NDMA and then the expression of several select proteins in the cell cytoplasmic and nuclear fractions were determined by Western blot analyses. The results indicated that NDMA enhanced expression of iNOS, phospho-PI3K, and phospho-IκBα in the cytoplasmic fraction of the PMN and PBMC. The nuclear fraction of these cells also had a higher NF-κB expression. Moreover, in PMN, NDMA caused an increased expression of phospho-Akt (T308), phospho-Akt (S473), and phospho-IKKαβ in the cytoplasm, and c-Jun and FosB in the nuclear fraction. Blocking of PI3K caused a decrease in expression of all these proteins in NDMA-exposed PMN. However, inhibition of PI3K led to a drop in expression of iNOS, phospho-PI3K, and phospho-IκBα in the cytoplasm, and in NF-κB in the nuclear fraction, of PBMC. The results of these studies indicated to us that NDMA activates the PI3K-Akt/PKB pathway in human PMN and that this, in turn, contributes to the activation of transcription factors NF-κB, c-Jun, and FosB involved in NO production (through modulation of iNOS expression). © 2014 Informa Healthcare USA, Inc. All rights reserved.


PubMed | Regional Center for Transfusion Medicine
Type: Journal Article | Journal: Journal of physiology and pharmacology : an official journal of the Polish Physiological Society | Year: 2015

Ozonated blood therapy is used in the treatment of several diseases, including superficial infections, burns, dental and intestinal conditions. Except that, the possibility of using ozone to sterilize blood supplies is under promising investigation. However, still little is known regarding the impact of blood ozonation, especially on biologically active serum sphinoglipids. In the present work we sought to investigate the contents of sphingolipids, such as sphingosine, sphingosine-1-phosphate (S-1-P), sphinganine, and ceramide (CER) in the plasma, after immediate and prolonged (1 h) ozonation of human whole blood. For the measurements liquid chromatography hyphenated with the mass spectrometry was applied. We demonstrated that only the content of sphingosine-1-phosphate in the plasma was increased significantly, possibly exerting its beneficial effect for various physiological and clinical events.

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