Maria Olszewska

Publications of Maria Olszewska

• Disrupted pro- and antioxidative balance as a mechanism of neurotoxicity induced by perinatal exposure to lead.

  Authors: Irena Baranowska-Bosiacka, Izabela Gutowska, Mariola Marchlewicz, Carla Marchetti, Mateusz Kurzawski, Viotetta Dziedziejko, Agnieszka Kolasa, Maria Olszewska, Marta Rybicka, Krzysztof Safranow, Przemysław Nowacki, Barbara Wiszniewska, Dariusz Chlubek

  Brain research. 12/2011; 1435:56-71.

  The aim of this paper was to examine if pre- and neonatal exposure that results in lead (Pb) concentration below 'safe level' (10 μg/dL) in offspring blood may cause disruption of the pro/antioxidantThe aim of this paper was to examine if pre- and neonatal exposure that results in lead (Pb) concentration below 'safe level' (10 μg/dL) in offspring blood may cause disruption of the pro/antioxidant balance in the developing rat brain. We studied oxidative stress intensity (malondialdehyde (MDA) concentration) as well as mRNA, protein expression and the activity of copper/zinc superoxide dismutase (SOD1), manganese superoxide dismutase (SOD2), glutathione peroxidase (GPx), phospholipid hydroperoxide glutathione peroxidase (GPx4), catalase (CAT), glutathione reductase (GSR). We also measured glutathione (GSH) concentrations in selected structures of the rat brain (forebrain cortex, FC, cerebellum, C, and hippocampus, H) and showed cellular localization of GPx4, SOD1 and SOD2 expressions in the hippocampus by immunohistochemical examinations. Despite low Pb level in blood we observed decrease of the activity of some antioxidant enzymes as well as mRNA and protein expression downregulation associated with an increase of MDA level and CAT expression upregulation, especially in the hippocampus region. At the subcellular level, downregulation of SOD2 expression and decreased enzyme activity as well as mitochondrial pool of GSH suggest also that mitochondrial mechanisms might account for Pb neurotoxicity mechanism. For some enzymes, we found differences in the effects of Pb on the level of expression and activity. The activity of CAT decreased despite an increase in mRNA and protein expression; and likewise the activities SOD1, GPx1 GPx4 decreased, despite substantially unchanged level of expression. These effects may be the result of impairment of catalytic function of the enzyme protein caused by Pb interaction or of reduction in the availability of cofactors. We conclude that antioxidant system of the hippocampus of immature rat brain is highly vulnerable to perinatal Pb exposure. Therefore, oxidative stress may be one of the possible mechanisms disturbing cellular metabolism in this structure. Disruption of pro- and antioxidant balance should be considered as a potential mechanism of the observed Pb adverse effects, leading to the impaired learning ability caused by Pb exposure in children.

• Presence of glucose in dialyzing fluid and synthesis of selected lipoxygenase-derived eicosanoids during hemodialysis.

  Authors: Barbara Dołęgowska, Wojciech Błogowski, Joanna Stępniewska, Krzysztof Safranow, Katarzyna Jakubowska, Maria Olszewska

  International urology and nephrology. 11/2011;

  PURPOSE: Active metabolites of arachidonic acid (AA), eicosanoids, are exerting a significant influence on renal homeostasis. In our recent paper, we demonstrated that high levels of lipoxygenasePURPOSE: Active metabolites of arachidonic acid (AA), eicosanoids, are exerting a significant influence on renal homeostasis. In our recent paper, we demonstrated that high levels of lipoxygenase (LOX)-derived AA metabolites, hydroxyeicosatetraenoic acids (HETEs), unfavorably influence post-transplant function of kidney allografts. Hence, in this study, we wanted (1) to analyze 5-, 12- and 15-HETE levels in non-diabetic chronic kidney disease (CKD) patients, already undergoing regular hemodialysis treatment, and determine factors that may influence these eicosanoids' generation, as well as, (2) to verify whether application of glucose-containing, instead of glucose-free, dialyzing fluids may be beneficial for the limitation of 5-, 12- and 15-HETE synthesis during a single hemodialysis session. METHODS: Twenty-four healthy individuals and 50 CKD patients undergoing regular hemodialysis treatment were included in the study. CKD patients were divided into two subgroups depending on presence/absence of glucose in dialyzing fluid. LOX-derived HETEs were measured using liquid chromatography. RESULTS: Results demonstrated higher levels of examined eicosanoids in CKD patients (P < 0.05 for all). Older age and higher C-reactive protein levels were associated with HETEs concentrations. Presence of glucose in dialyzing fluid significantly diminished the increase in 5- and 12-HETE synthesis (411.24% vs. 107.29%, P < 0.006; 301.70% vs. 98.21%, P < 0.0008, respectively), however, it did not influence 15-HETE generation (156.98% vs. 135.24%, P = 0.26). CONCLUSIONS: Non-diabetic CKD patients have higher levels of LOX-derived HETEs, which are associated mainly with age and intensified inflammatory process. The presence of glucose in the dialysate is associated with a reduced synthesis of selected LOX-derived HETEs in these patients; hence, we speculate that the application of such a simple hemodialysis fluid modification may have a favorable influence on post-transplant outcomes.

• Polymorphism of the CD36 Gene and Cardiovascular Risk Factors in Patients with Coronary Artery Disease Manifested at a Young Age.

  Authors: Monika Ewa Rać, Janina Suchy, Grzegorz Kurzawski, Agnieszka Kurlapska, Krzysztof Safranow, Michał Rać, Dagmara Sagasz-Tysiewicz, Andrzej Krzystolik, Wojciech Poncyljusz, Katarzyna Jakubowska, Maria Olszewska, Beata Krupa, Dariusz Chlubek

  Biochemical genetics. 11/2011; 50(1-2):103-11.

  This study investigates potential associations between CD36 gene variants and the presence of risk factors in Caucasians with coronary artery disease (CAD) manifested at a young age. The study groupThis study investigates potential associations between CD36 gene variants and the presence of risk factors in Caucasians with coronary artery disease (CAD) manifested at a young age. The study group consisted of 90 patients; the men were ≤ 50 years old and the women were ≤ 55 years old. Amplicons of exons 4 and 5 including fragments of introns were analyzed by DHPLC. Two polymorphisms were found: IVS3-6 T/C (rs3173798) and IVS4-10 G/A (rs3211892). The C allele of the IVS3-6 T/C polymorphism was associated with higher prevalence of obesity and diabetes, higher hsCRP, lower Lp(a) serum concentrations, and younger age at myocardial infarction. The A allele of the IVS4-10 G/A polymorphism was associated with older age of myocardial infarction and higher white blood cell count. The functional role of CD36 polymorphisms in CAD development needs further research.

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• AMPD1 gene mutations are associated with obesity and diabetes in Polish patients with cardiovascular diseases.

  Authors: Krzysztof Safranow, Janina Suchy, Katarzyna Jakubowska, Maria Olszewska, Agnieszka Bińczak-Kuleta, Grzegorz Kurzawski, Ryszard Rzeuski, Edyta Czyżycka, Beata Łoniewska, Zdzisława Kornacewicz-Jach, Andrzej Ciechanowicz, Dariusz Chlubek

  Journal of applied genetics. 02/2011; 52(1):67-76.

  Previous studies showed an association of the common functional polymorphism (C34T, Gln12Stop) in the adenosine monophosphate deaminase-1 (AMPD1) gene with survival in heart failure (HF) and/orPrevious studies showed an association of the common functional polymorphism (C34T, Gln12Stop) in the adenosine monophosphate deaminase-1 (AMPD1) gene with survival in heart failure (HF) and/or coronary artery disease (CAD). The aim of the study was to search for other mutations in selected regions of the AMPD1 gene in Polish CAD and HF patients, and to analyze their associations with obesity and diabetes. Exons 2, 3, 5, and 7 of AMPD1 were scanned for mutations in 97 patients with CAD without HF (CAD+ HF-), 104 patients with HF (HF+), and 200 newborns from North-Western Poland using denaturing high-performance liquid chromatography (DHPLC), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and direct sequencing. Frequencies of AMPD1 C34T mutation, as well as novel A99G, G512A, IVS4-6delT, and C784T sequence alterations, were similar in the three groups, but 860T mutated allele was less frequent in the combined CAD+ HF- and HF+ groups than in the controls (1.7% vs. 4.3%, p=0.040). Heterozygous 34CT genotype was associated with lower (odds ratio [OR]=0.32, 95% confidence interval [CI] =0.13-0.81) and 860AT with higher (OR=13.7, 95%CI=1.6-118) prevalence of diabetes or hyperglycemia in relation to wild-type homozygotes. Abdominal obesity was more frequent in 860AT patients than in wild-type homozygotes and 34CT heterozygotes (86% vs. 40% vs. 29%, p<0.05). Nine genes containing polymorphisms linked with AMPD1 C34T mutation were found in the HapMap database. AMPD1 C34T nonsense mutation is associated with reduced prevalence of diabetes and obesity in patients with CAD or HF, but A860T substitution seems to exert opposite metabolic effects and should always be accounted for in the studies of the AMPD1 genotype.

• The effect of L-ascorbic acid and/or tocopherol supplementation on electrophysiological parameters of the colon of rats chronically exposed to lead.

  Authors: Danuta I Kosik-Bogacka, Irena Baranowska-Bosiacka, Mariola Marchlewicz, Agnieszka Kolasa, Katarzyna Jakubowska, Maria Olszewska, Natalia Łanocha, Ireneusz Wiernicki, Barbara Millo, Barbara Wiszniewska, Dariusz Chlubek

  Medical science monitor : international medical journal of experimental and clinical research. 01/2011; 17(1):BR16-26.

  The aim of this study was to assess the effect of diet supplementation with L-ascorbic acid (500 mg/L), tocopherol (3 mg/kg b.w.), and/or a water soluble analog of tocopherol (Trolox) (48 mg/L) onThe aim of this study was to assess the effect of diet supplementation with L-ascorbic acid (500 mg/L), tocopherol (3 mg/kg b.w.), and/or a water soluble analog of tocopherol (Trolox) (48 mg/L) on ion transport in the colon of rats subjected to a chronic exposure (9 months) to 0.1% lead acetate in drinking water. The electrophysiological parameters of the colon wall were measured with Ussing methods. Lead content in the whole blood was analyzed by graphite furnace atomic absorption spectrometry (GFAAS) using Zeeman correction. L-ascorbic acid and tocopherol in plasma was measured by high performance liquid chromatography. Immunohistochemical reaction was carried out for visualization of occludin, the intracellular tight junction protein. We showed a strong inhibitory effect of lead on the electrophysiological parameters, changes in intestinal permeability, disappearance of junctional occludin, decreased amount of mucus covering the colon surface, and the accumulation of PAS-positive substance in the apical region of the cytoplasm in the absorptive cells. Supplementation with tocopherol or Trolox did not exert a beneficial influence on the studied parameters. L-ascorbic acid positively influenced the examined electrophysiological parameters, as it cancelled the inhibitory influence of lead on ion transport in the rat colon. L-ascorbic acid also protected against tight junction disruption of epithelial cells in the colon of the lead-treated rats. A similar effect was observed in the group of rats receiving lead and supplemented with L-ascorbic acid plus Trolox.

• [Metformin - mechanisms of action and use for the treatment of type 2 diabetes mellitus].

  Authors: Marzena Grzybowska, Joanna Bober, Maria Olszewska

  Postȩpy higieny i medycyny doświadczalnej (Online). 01/2011; 65:277-85.

  Metformin is widely used for the treatment of type 2 diabetes mellitus. Although this biguanide derivative has been used for more than 50 years, its mechanism of action has not been fully elucidated.Metformin is widely used for the treatment of type 2 diabetes mellitus. Although this biguanide derivative has been used for more than 50 years, its mechanism of action has not been fully elucidated. In this article we describe the latest achievements concerning the mechanisms of antihyperglycemic action of metformin. They include: decrease of glucose absorption in the small intestine, increase of glucose transport into cells, decrease in the plasma free fatty acid concentrations and inhibition of gluconeogenesis. Activation of AMP-activated protein kinase (AMPK) plays an important role in these processes. The latest discoveries have revealed mechanisms of anti-atherosclerotic, hypotensive and anticancer action of metformin and its impact on vein endothelial function. The pleiotropic actions of metformin include impact on plasma lipid profile, decrease of oxidative stress, and increase in plasma fibrinolytic activity. Although metformin is not metabolized, the latest research has shown that it is actively transported into hepatocytes and renal tubular epithelium, by OCT1 (organic cation transporter 1, encoded by the SLC22A1 gene) and OCT2 (organic cation transporter 2, encoded by the SLC22A2 gene), respectively. However, MATE1 transporter (multidrug and toxin extrusion 1 protein) is encoded by the SLC47A1 gene and facilitates metformin excretion from these cells into bile and urine. Metformin transporter gene polymorphisms may contribute to significant variation in drug response. Further studies of mechanisms of metformin action could contribute to its wider use for the prevention of type 2 diabetes mellitus, cancer, and Alzheimer’s disease, and for the treatment of type 1 diabetes mellitus, and polycystic ovary syndrome (PCOS).

• Lipoxygenase-derived hydroxyeicosatetraenoic acids--novel perioperative markers of early post-transplant allograft function?

  Authors: Barbara Dolegowska, Wojciech Blogowski, Krzysztof Safranow, Leszek Domanski, Katarzyna Jakubowska, Maria Olszewska

  Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association. 12/2010; 25(12):4061-7.

  Active metabolites of arachidonic acid (AA), eicosanoids, strongly influence renal homeostasis. The aims of this study were to measure perioperative variations in lipoxygenase (LOX)-derived 5-, 12-Active metabolites of arachidonic acid (AA), eicosanoids, strongly influence renal homeostasis. The aims of this study were to measure perioperative variations in lipoxygenase (LOX)-derived 5-, 12- and 15-hydroxyeicosatetraenoic (HETE) acids levels, and to examine whether (i) dynamics of these eicosanoid generation changes during the first 5 min of renal allograft reperfusion, (ii) examined HETE acids may influence perioperative 20-HETE generation, and (iii) LOX HETE may serve as perioperative markers of early post-transplant allograft function. Sixty-nine kidney recipients were divided into early, slow and delayed graft function (EGF, SGF and DGF, respectively) groups. Blood was taken directly before, and in the consecutive minutes of graft reperfusion. HETE concentrations were measured using liquid chromatography. Creatinine levels were measured during the perioperative period, as well as during follow-up visits (first post-transplant year). Our results demonstrated significant differences in the concentrations and dynamics of HETE changes between the examined groups. Moreover, observed changes in HETE concentrations were strongly associated with post-transplant graft function and perioperative 20-HETE synthesis. Application of cut-off limits for newly introduced markers, that is 71.72 ng/mL for 5-HETE(5), 12.3 ng/mL for 12-HETE△(5-0) and -6.1 ng/mL for 15-HETE△(5-0), resulted in 72.5-81.5% sensitivity and 50-54% specificity for SGF/DGF prediction. Moreover, mixed model analysis revealed that recipients classified according to results of 5-HETE(5) and 15-HETE△(5-0) significantly differ in 1-year post-transplant allograft function (P  =  0.03 and P  < 0.05, respectively), however, not in the frequency of acute rejections' episodes (P = 0.91 and P = 0.31, respectively). We hereby report that human kidney transplantations are accompanied by significant changes in LOX AA metabolism, which strongly influences and predicts early (1 year) post-transplant graft function.

• [Xanthinuria type I as the cause of nephrolithiasis in 17-years old girl].

  Authors: Barbara Borucka, Dariusz Runowski, Krzysztof Safranow, Maria Olszewska, Katarzyna Jakubowska, Dariusz Chlubek

  Polski merkuriusz lekarski : organ Polskiego Towarzystwa Lekarskiego. 08/2010; 29(170):111-4.

  Classic xanthinuria is a rare metabolic defect concerning the final reactions of purine catabolism. There are two types of the disorder: type I results from xanthine dehydrogenase (XDH) deficiency,Classic xanthinuria is a rare metabolic defect concerning the final reactions of purine catabolism. There are two types of the disorder: type I results from xanthine dehydrogenase (XDH) deficiency, while type II is characterized by lack of both XDH and aldehyde oxidase activity. Both types are clinically similar and are characterized by elevated xanthine concentration in body fluids that can lead to xanthine crystallisation. The most common manifestation of the disease is urolithiasis, but in most cases xanthinuria remains asymptomatic and the diagnosis is accidental. In the paper we report the first case study of xanthinuria in Poland in a child presenting with urolithiasis. 17-years old female patient was diagnosed because of recurrent urinary lithiasis and hypouricemia was detected during routine tests. Plasma and urine concentrations of oxypurines were measured by high-performance liquid chromatography (HPLC) and showed typical features of xanthinuria: hypouricemia, hypouricosuria, xanthinuria and elevated plasma xanthine. The allopurinol loading test demonstrated type I xanthinuria. The presented case report supports that first symptoms of xanthinuria can appear at any age and this disorder should be considered during diagnosing urolithiasis.

• Analysis of human CD36 gene sequence alterations in the oxidized low-density lipoprotein-binding region using denaturing high-performance liquid chromatography.

  Authors: Monika Ewa Rać, Janina Suchy, Grzegorz Kurzawski, Krzysztof Safranow, Katarzyna Jakubowska, Maria Olszewska, Barbara Garanty-Bogacka, Michał Rać, Wojciech Poncyljusz, Dariusz Chlubek

  Genetic testing and molecular biomarkers. 08/2010; 14(4):551-7.

  Denaturing high-performance liquid chromatography (DHPLC) has been employed as a prescreening tool to reduce the amount of DNA sequencing. It could be a simple and cost-effective screening method forDenaturing high-performance liquid chromatography (DHPLC) has been employed as a prescreening tool to reduce the amount of DNA sequencing. It could be a simple and cost-effective screening method for mutations and polymorphisms in exons 4, 5, and 6 of the CD36 gene, which encode the protein region responsible for the removal of oxidized low-density lipoprotein. Genomic DNA was isolated from 306 Caucasian infants of Polish origin. Six single-nucleotide substitutions were detected by DHPLC and confirmed by direct sequencing. The A591T, G550A, and C572T alterations have not been described so far. Each of two nonsynonymous substitutions (Asp184Asn, Pro191Leu) was found in one subject (0.2% minor allele frequency). The results suggest that nonsynonymous alterations in the analyzed CD36 region are rare in Caucasians. DHPLC is a specific and cost-effective technique that may prove to be particularly useful for the identification of polymorphisms and mutations in the CD36 gene.

• Microelements in Stones, Urine, and Hair of Stone Formers: A New Key to the Puzzle of Lithogenesis?

  Authors: Marcin Słojewski, Bogusław Czerny, Krzysztof Safranow, Katarzyna Jakubowska, Maria Olszewska, Andrzej Pawlik, Adam Gołąb, Marek Droździk, Dariusz Chlubek, Andrzej Sikorski

  Biological trace element research. 12/2009;

  The role of trace elements in lithogenesis is still unclear. The aim of this study was to evaluate the levels of elements in urinary stones and in the urine and hair of stone formers to identifyThe role of trace elements in lithogenesis is still unclear. The aim of this study was to evaluate the levels of elements in urinary stones and in the urine and hair of stone formers to identify these elements that have synergic correlations in studied materials and may contribute to lithogenesis. A total of 219 consecutive patients with idiopathic upper urinary tract stones were prospectively enrolled in the study. Urine and hair samples were collected from all patients. The content of the stone was evaluated using atomic absorption spectrometry, spectrophotometry, and colorimetric methods. The analysis of 29 elements in stones and hair and 21 elements in urine was performed using inductively coupled plasma-atomic emission spectrometry. The strength of correlation was described with the value of Spearman's rank correlation coefficient. The positive correlation between concentration of sodium, potassium, magnesium, barium, vanadium, zinc, silicon, phosphorus, and iodine in phosphate stones was observed. Only a few incidental correlations between the composition of stones and the distribution of elements in urine and in hair were found. There were 109 positive two-element correlations between two materials. The most common were observed for vanadium, aluminum, lead, cobalt, and molybdenum. Two-element positive correlations for all samples were established only for three elements: vanadium, lead, and aluminum. Results indicate that analysis of particular elements in hair and urine cannot predict the composition of urinary stones. This study showed, for the first time, correlations between the levels of vanadium, lead, and aluminum in the stones, urine, and hair of stone formers.

• PIN1 gene variants in Alzheimer's disease.

  Authors: Aleksandra Maruszak, Krzysztof Safranow, Katarzyna Gustaw, Beata Kijanowska-Haladyna, Katarzyna Jakubowska, Maria Olszewska, Maria Styczynska, Mariusz Berdynski, Andrzej Tysarowski, Dariusz Chlubek, Janusz Siedlecki, Maria Barcikowska, Cezary Zekanowski

  BMC medical genetics. 11/2009; 10(1):115.

  ABSTRACT: BACKGROUND: Peptidyl-prolyl isomerase, NIMA-interacting 1 (PIN1) plays a significant role in the brain and is implicated in numerous cellular processes related to Alzheimer's disease (AD)ABSTRACT: BACKGROUND: Peptidyl-prolyl isomerase, NIMA-interacting 1 (PIN1) plays a significant role in the brain and is implicated in numerous cellular processes related to Alzheimer's disease (AD) and other neurodegenerative conditions. There are confounding results concerning PIN1 activity in AD brains. Also PIN1 genetic variation was inconsistently associated with AD risk. METHODS: We performed analysis of coding and promoter regions of PIN1 in early- and late-onset AD and frontotemporal dementia (FTD) patients in comparison with healthy controls. RESULTS: Analysis of eighteen PIN1 common polymorphisms and their haplotypes in EOAD, LOAD and FTD individuals in comparison with the control group did not reveal their contribution to disease risk. In six unrelated familial AD patients four novel PIN1 sequence variants were detected. c.58+64C>T substitution that was identified in three patients, was located in an alternative exon. In silico analysis suggested that this variant highly increases a potential affinity for a splicing factor and introduces two intronic splicing enhancers. In the peripheral leukocytes of one living patient carrying the variant, a 2.82 fold decrease in PIN1 expression was observed. CONCLUSIONS: Our data does not support the role of PIN1 common polymorphisms as AD risk factor. However, we suggest that the identified rare sequence variants could be directly connected with AD pathology, influencing PIN1 splicing and/or expression.

• Does smoking have any effect on urinary stone composition and the distribution of trace elements in urine and stones?

  Authors: Marcin Słojewski, Bogusław Czerny, Krzysztof Safranow, Marek Droździk, Andrzej Pawlik, Katarzyna Jakubowska, Maria Olszewska, Adam Gołąb, Elżbieta Byra, Dariusz Chlubek, Andrzej Sikorski

  Urological research. 10/2009;

  The role of particular elements in lithogenesis is still unclear and debated. Probably some of them may promote or conversely inhibit crystal nucleation of organic or mineral species. A fewThe role of particular elements in lithogenesis is still unclear and debated. Probably some of them may promote or conversely inhibit crystal nucleation of organic or mineral species. A few epidemiological data link smoking with the risk of calcium stones. The aim of this hospital-based study was to evaluate the distribution of trace elements in urine and urinary stones, and possible correlation with stone constituents in smoking and non-smoking individuals. 209 stones and urine samples collected from idiopathic stone-formers were analyzed to evaluate the mineral composition and the distribution of elements, 29 in stones and 21 in urine. Values were statistically compared considering smoking, arterial hypertension and coronary heart disease as grouping variables. No differences were noted either for comparison of mineral components or the elements concentrations in stones in both groups. The concentration of mercury in urine was higher in smokers than in non-smokers, but the statistical significance was at the moderate level. Our findings do not support the concept of possible association between smoking and urinary lithogenesis, but we believe that further investigations are needed in this area.

• Platelets arachidonic acid metabolism in patients with essential hypertension.

  Authors: Barbara Dołegowska, Wojciech Błogowski, Karolina Kedzierska, Krzysztof Safranow, Katarzyna Jakubowska, Maria Olszewska, Monika Rać, Dariusz Chlubek, Kazimierz Ciechanowski

  Platelets. 07/2009; 20(4):242-9.

  Arachidonic acid's (AA) metabolites, eicosanoids, exert a tremendous influence on circulatory and vascular homeostasis, and in humans are generated by many organs and cell types. In this study weArachidonic acid's (AA) metabolites, eicosanoids, exert a tremendous influence on circulatory and vascular homeostasis, and in humans are generated by many organs and cell types. In this study we wanted to verify whether platelets AA metabolism play a significant role in pathogenesis of essential hypertension (EH). Participants were divided into the study (EH) and the control group. Plasma and urine concentrations of isoprostanes (8-iPF(2alpha)-III) and thromboxane B(2) (TxB(2)) were determined using the ELISA method. The levels of 5- and 12-hydroxyeicosatetraenoic (HETE) acids, generated by platelets, were analysed using RP-HPLC. In a suspension of not stimulated and AA-stimulated platelets TxB(2) level was statistically lower in the study than in the control group (p < 0.0001 and 0.001 respectively). The concentration of 12-HETE was significantly elevated in EH patients compared to the control group; however, only in the non-stimulated conditions (p < 0.05). Plasma and urine F2-isoprostanes levels were significantly higher in hypertensive individuals than in the control group (p < 0.00002 and p < 0.01 respectively). Moreover, EH patients excreted more TxB(2) in urine than normotensive individuals (p < 0.05). Our results highlight the mutual connections between the platelets AA metabolism and indicate its possible role in the pathogenesis of arterial hypertension. Moreover, we hypothesize that platelets AA metabolism may exert a pro-atherosclerotic effect. Finally, we suggest the use of (5-HETE+12-HETE)/TxB(2) parameter in further studies.

• PIN1 gene variants in Alzheimer's disease

  Authors: Aleksandra Maruszak, Krzysztof Safranow, Katarzyna Gustaw, Beata Kijanowska-Haładyna, Katarzyna Jakubowska, Maria Olszewska, Maria Styczyńska, Mariusz Berdyński, Andrzej Tysarowski, Dariusz Chlubek, Janusz Siedlecki, Maria Barcikowska, Cezary Żekanowski

  BMC Medical Genetics. 01/2009;

  Abstract Background Peptidyl-prolyl isomerase, NIMA-interacting 1 (PIN1) plays a significant role in the brain and is implicated in numerous cellular processes related to Alzheimer's disease (AD)Abstract Background Peptidyl-prolyl isomerase, NIMA-interacting 1 (PIN1) plays a significant role in the brain and is implicated in numerous cellular processes related to Alzheimer's disease (AD) and other neurodegenerative conditions. There are confounding results concerning PIN1 activity in AD brains. Also PIN1 genetic variation was inconsistently associated with AD risk. Methods We performed analysis of coding and promoter regions of PIN1 in early- and late-onset AD and frontotemporal dementia (FTD) patients in comparison with healthy controls. Results Analysis of eighteen PIN1 common polymorphisms and their haplotypes in EOAD, LOAD and FTD individuals in comparison with the control group did not reveal their contribution to disease risk. In six unrelated familial AD patients four novel PIN1 sequence variants were detected. c.58+64C>T substitution that was identified in three patients, was located in an alternative exon. In silico analysis suggested that this variant highly increases a potential affinity for a splicing factor and introduces two intronic splicing enhancers. In the peripheral leukocytes of one living patient carrying the variant, a 2.82 fold decrease in PIN1 expression was observed. Conclusion Our data does not support the role of PIN1 common polymorphisms as AD risk factor. However, we suggest that the identified rare sequence variants could be directly connected with AD pathology, influencing PIN1 splicing and/or expression.

• Association of C34T AMPD1 gene polymorphism with features of metabolic syndrome in patients with coronary artery disease or heart failure.

  Authors: Krzysztof Safranow, Edyta Czyzycka, Agnieszka Binczak-Kuleta, Ryszard Rzeuski, Janusz Skowronek, Andrzej Wojtarowicz, Katarzyna Jakubowska, Maria Olszewska, Beata Loniewska, Robert Kaliszczak, Zdzislawa Kornacewicz-Jach, Andrzej Ciechanowicz, Dariusz Chlubek

  Scandinavian journal of clinical and laboratory investigation. 10/2008;

  Objective. The common C34T polymorphism in the AMP deaminase-1 (AMPD1) gene results in an inactive enzyme in homozygotes for the mutated T allele. Some studies have shown an association of T alleleObjective. The common C34T polymorphism in the AMP deaminase-1 (AMPD1) gene results in an inactive enzyme in homozygotes for the mutated T allele. Some studies have shown an association of T allele with longer survival in heart failure (HF) and/or coronary artery disease (CAD). The aim of this study was to assess genotype-phenotype correlations in such patients, with emphasis on components of the metabolic syndrome. Methods. Ninety-seven patients with CAD without HF (CAD+ HF-) and 104 with HF (HF+) were genotyped by PCR-RFLP. The genetic control group comprised 200 newborns. Results. No significant differences were found in the frequency of AMPD1 genotypes between the groups. In the CAD+ HF- group, the carriers of T allele compared to CC homozygotes had significantly lower values of waist circumference (89.5+/-8.5 versus 97.7+/-11.2 cm; p = 0.00029), waist/hip ratio (p = 0.0059) and BMI (p = 0.045). There was no diabetes or fasting glycaemia >/=126 mg/dL in T carriers, while these features were present in 25 % of CC homozygotes (p = 0.0024). In the HF+ group, a tendency towards a lower prevalence of diabetes (20 % versus 41 %; p = 0.068) and significantly lower systolic blood pressure (p = 0.048) were observed in T allele carriers. Conclusions. C34T AMPD1 polymorphism may be associated with reduced frequency of obesity in CAD patients and of hyperglycaemia and diabetes in both CAD and HF patients. Morphometric parameters associated with adipose tissue distribution and parameters of glucose metabolism should be analysed as potential confounders in further studies on the role of polymorphisms of AMPD1 and other genes associated with AMP and adenosine metabolism in cardiovascular disease.

• The effect of preservation solutions UW and EC on purine concentration in rat kidney.

  Authors: Leszek Domański, Tadeusz Sulikowski, Maciej Romanowski, Krzysztof Safranow, Andrzej Pawlik, Katarzyna Jakubowska, Violetta Dziedziejko, Magda Wiśniewska, Maciej Domański, Dariusz Chlubek, Maria Olszewska, Kazimierz Ciechanowski

  Annales Academiae Medicae Stetinensis. 02/2008; 54(1):53-9.

  INTRODUCTION: Ischemia/reperfusion injury in organ transplantation is a multifactor process that may lead to organ damage and primary graft dysfunction. Perfusion is a process which creates aINTRODUCTION: Ischemia/reperfusion injury in organ transplantation is a multifactor process that may lead to organ damage and primary graft dysfunction. Perfusion is a process which creates a possibility of graft injury. Preservation solutions are thought to diminish the ischemic injury and an appropriate choice of the solution should guarantee a better graft function and good prognosis for graft survival. The aim of the study was to examine the effect of preservation solutions University of Wisconsin (UW) and Euro-Collins (EC) on purine concentration in rat kidney. MATERIAL AND METHODS: The study was carried out on Wistar rat kidneys divided into 3 groups: kidneys perfused with 0.9% NaCl (control group), kidneys perfused with UW preservation solution, kidneys perfused with EC preservation solution. After bilateral nephrectomy the right kidney was immediately frozen in liquid nitrogen and the left kidney was placed in 4 degrees C UW, EC or 0.9% NaCl solution for 24 hours. The concentrations of purine nucleotides were determined using high-performance liquid chromatography (HPLC) method. RESULTS: The concentrations ofpurine nucleotides in renal tissue without cold ischemia did not differ significantly between rats perfused with 0.9% NaCl, UW and EC solutions. The tissue concentrations of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), total adenine nucleotides (TAN), guanosine monophosphate (GMP) and inosine monophosphate (IMP) were significantly increased, whereas the concentrations of adenosine (Ado), inosine (Ino), guanosine (Guo), hypoxanthine (Hyp) and xanthine (Xan) were significantly lower in rats perfused with EC solution in comparison to rats perfused with 0.9% NaCl. CONCLUSIONS: Purine concentration profile in rat kidney reflects a protective influence of EC and UW solutions on high-energy nucleotides in conditions when their depletion might be harmful to graft function.

• [Oxidative stress and trace elements affect the activity of sodium transporting systems in a cellular membrane of a erythrocyte]

  Authors: Karolina Kedzierska, Joanna Bober, Ewa Kwiatkowska, Ewa Stachowska, Maria Olszewska, Dariusz Chlubek, Kazimierz Ciechanowski

  Annales Academiae Medicae Stetinensis. 01/2008; 54(3):105-14.

  We examined the activity of sodium transporting systems (STS) in a cellular membrane of erythrocytes in a group of 21 patients under chronic hemodialysis treatment with the dialyzing fluid containingWe examined the activity of sodium transporting systems (STS) in a cellular membrane of erythrocytes in a group of 21 patients under chronic hemodialysis treatment with the dialyzing fluid containing glucose HD-g(+), and 22 patients dialysed with the fluid not containing glucose HD-g(-), 21 patients with chronic kidney failure already not treated with dialysis and 21 control group. We examined the concentration of antioxidative system cofactors, such as zinc, copper and selenium in erythrocytes and plasma. The marker of oxidative stress in erythrocytes and plasma was the concentration of TBARS. Among all STS we examined the activity of Na(+)/K(+) ATP-ase, Na(+)/K(+)/Cl(-); co-transport, Na(+)/Li(+) exchanger, Na(+), K(+)-outflow. Copper zinc and selenium as cofactors of antioxidative enzymes may reflect the antioxidative processes inside the organism undergoing the influence of free radicals.

• Effect of conjugated linoleic acids on the activity and mRNA expression of 5- and 15-lipoxygenases in human macrophages.

  Authors: Ewa Stachowska, Violetta Dziedziejko, Krzysztof Safranow, Katarzyna Jakubowska, Maria Olszewska, Bogusław Machaliñski, Dariusz Chlubek

  Journal of agricultural and food chemistry. 06/2007; 55(13):5335-42.

  Lipoxygenases are a family of non-heme enzyme dioxygenases. The role of lipoxygenases is synthesis of hydroperoxides of fatty acids, which perform signaling functions in the body. Studies onLipoxygenases are a family of non-heme enzyme dioxygenases. The role of lipoxygenases is synthesis of hydroperoxides of fatty acids, which perform signaling functions in the body. Studies on conjugated linoleic acids (CLAs) as fatty acids with a potential anti-atherosclerotic function have recently been initiated. The aim of the study was to test the effect of CLAs and linoleic acid on 5- and 15-lipoxygenase (5-LO, 15-LO-1) enzyme activity, their mRNA expression, and concentration in the cells. It was also desired to determine whether the CLAs are substrates for the enzymes. For the experiments monocytic cell line (THP-1) and monocytes obtained from human venous blood were used. Monocytes were differentiated to macrophages: THP-1 (CD14+) by PMA administration (100 nM for 24 h) and monocytes from blood (CD14+) by 7-day cultivation with the autologous serum (10%). After differentiation, macrophages were cultured with 30 microM CLAs or linoleic acid for 48 h. The 15- and 5-lipoxygenase products were measured by HPLC method. mRNA expression and protein content were analyzed by real-time PCR and Western blot analysis. The in vitro studies proved that both CLA isomers are not substrates for 15-LO-1; in ex vivo studies hydroxydecadienoic acid (HODE) concentration was significantly reduced (p = 0.019). The trans-10,cis-12 CLA isomer reduced HODE concentration by 28% (p = 0.046) and the cis-9,trans-11 CLA isomer by 35% (p = 0.028). In macrophages obtained from THP-1 fatty acids did not change significantly mRNA expression of the majority of the investigated genes. CLAs did not change the content of 5-LO and 15-LO-1 proteins in macrophages obtained from peripheral blood. Linoleic acid induced 15-LO-1 expression (2.6 times, p < 0.05). CLAs may perform the function of an inhibitor of lipoxygenase 15-LO-1 activity in macrophages.

• Influence of glucose in the dialysate on the activity of erythrocyte-glutathione-peroxidase and blood selenium concentration in hemodialyzed patients.

  Authors: Joanna Bober, Ewa Kwiatkowska, Karolina Kedzierska, Maria Olszewska, Edyta Gołebiewska, Ewa Stachowska, Elzbieta Kucharska, Kazimierz Ciechanowski, Dariusz Chlubek

  Archives of medical research. 05/2007; 38(3):330-6.

  BACKGROUND: The aim of the study was to show the influence of glucose in the dialysate on the intensity of oxidative stress, activity of glutathione peroxidase (GSHPx) and concentration of seleniumBACKGROUND: The aim of the study was to show the influence of glucose in the dialysate on the intensity of oxidative stress, activity of glutathione peroxidase (GSHPx) and concentration of selenium in patients undergoing regular hemodialysis. METHODS: The study was comprised of 85 patients hemodialyzed with dialysate containing glucose [HD-g(+)] or not containing glucose [HD-g(-)], patients with chronic renal failure on conservative treatment and control group. The concentrations of the products of reaction with thiobarbituric acid (TBARS), concentration of selenium in erythrocytes and plasma, concentration of copper in erythrocytes and the activity of GSHPx were determined. RESULTS: GSHPx had significantly higher activity in HD-g(-) group before HD than in control group. In HD-g(+) group before hemodialysis, the activity of GSHPx was significantly lower than in the control group. After HD, the activity showed a statistically significant increase. In both hemodialyzed groups, selenium concentration before hemodialysis both in plasma and erythrocytes was significantly lower, compared to control group. In the group of patients with CRF on conservative treatment, selenium concentration in RBC was significantly higher, compared to concentrations obtained in other groups except for control group. The increase of copper concentration in erythrocytes was accompanied by the increase of oxidative stress and increase of TBARS concentration. The opposite relationship was observed for selenium-its concentration was inversely correlated to copper concentration. CONCLUSIONS: In both groups of hemodialyzed patients, hemodialysis caused the increase of GSHPx in erythrocyte activity and increase of plasma and erythrocyte selenium concentration.

• Oxypurine and purine nucleoside concentrations in renal vein of allograft are potential markers of energy status of renal tissue.

  Authors: Leszek Domański, Krzysztof Safranow, Marek Ostrowski, Andrzej Pawlik, Maria Olszewska, Grazyna Dutkiewicz, Kazimierz Ciechanowski

  Archives of medical research. 03/2007; 38(2):240-6.

  BACKGROUND: Impairment of organ function derived from ischemia-reperfusion injury is an important problem in solid organ transplantation. Cell alterations induced by ischemia prime the tissue forBACKGROUND: Impairment of organ function derived from ischemia-reperfusion injury is an important problem in solid organ transplantation. Cell alterations induced by ischemia prime the tissue for subsequent damage that occurs during the reperfusion phase. Purine nucleosides and oxypurines are products of adenine nucleotides degradation. Reperfusion and reoxygenation are accompanied by production of reactive oxygen species and free radicals, which lead to damage of graft tissue. The aim of this study was to measure concentrations of adenine nucleotides and their metabolites in renal allograft vein as well as in recipient's peripheral veins during the reperfusion period and to evaluate their usefulness as markers of tissue metabolism in kidney allografts. METHODS: The study enrolled 20 renal transplant recipients. The first blood sample was taken from the recipient's ulnar vein before anastomosing of the kidney graft's vessels with recipient's iliac vessels. Samples were then taken from the renal allograft and ulnar veins 5 min after total graft reperfusion measured with an infrared camera. High-performance liquid chromatography (HPLC) was performed to measure whole blood and plasma concentrations of adenosine triphosphate (ATP), adenosine monophosphate (AMP), guanosine (Guo), inosine (Ino), hypoxanthine (Hyp), xanthine (Xan), uric acid (UA), and uridine (Urd). RESULTS: Hyp and Xan concentrations were significantly increased in renal allograft vein after reperfusion as compared with peripheral vein during the pre- and post-reperfusion periods. CONCLUSIONS: The results of the present study suggest that differences in Hyp and Xan concentrations between renal and peripheral veins reflect metabolic alterations in renal tissue during reperfusion and may be useful for graft function monitoring during reperfusion.