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A sensitive and specific liquid chromatography tandem mass spectrometry method for quantification of 1-methylpyridinium (1-MP) and 1,4-dimethylpyridinium (1,4-DMP) in rat plasma and tissues homogenates was developed. Chromatographic separation was performed on an Aquasil C18 analytical column with an isocratic elution of acetonitrile and water, both with an addition of formic acid (0.1%, v/v). Detection was achieved by triple quadrupole mass spectrometer TSQ Quantum Ultra equipped with a heated electrospray ionization source (HESI). The limit of quantification for both compounds was 0.05 pg/mL in plasma and 0.25 μg/g in studied tissues. The method was applied to pharmacokinetics and bioavailability of both 1-MP and 1,4-DMP with tissue distribution of 1,4-DMP in rats. Pharmacokinetic studies of 1-MP and 1,4-DMP were carried out following their intravenous or intragastric administration to male Wistar rats at the dose of 100 mg/kg. The terminal half-lives of I-MP and 1,4-DMP after their intravenous administration were 55.3 and 70.8 min, respectively. The absolute bioavailability was 51 and 31% for t-MP and 1,4-DMP, respectively.

Ambidexterity wyrażająca się w godzeniu napięć pomiędzy eksploracją i eksploatacją jest stosunkowo nowym problemem badawczym w naukach o zarządzaniu. Celem opracowania jest identyfikacja trendów badawczych nad tym zjawiskiem przy wykorzystaniu metodyki systematycznego przeglądu literatury. Na podstawie liczby publikacji i analizy ich treści określono aktywność badawczą w tym obszarze na przestrzeni czasu oraz wskazano główne perspektywy teoretyczne i poziomy analizy radzenia sobie z tego rodzaju napięciem.

1-Methylnicotinamide (MNA), the major endogenous metabolite of nicotinic acid (NicA), may partially contribute to the vasoprotective properties of NicA. Here we compared the antiatherosclerotic effects of MNA and NicA in apolipoprotein E (ApoE)/low-density lipoprotein receptor (LDLR)-deficient mice. ApoE/LDLR(-/-) mice were treated with MNA or NicA (100 mg/kg). Plaque size, macrophages, and cholesterol content in the brachiocephalic artery, endothelial function in the aorta, systemic inflammation, platelet activation, as well as the concentration of MNA and its metabolites in plasma and urine were measured. MNA and NicA reduced atherosclerotic plaque area, plaque inflammation, and cholesterol content in the brachiocephalic artery. The antiatherosclerotic actions of MNA and NicA were associated with improved endothelial function, as evidenced by a higher concentration of 6-keto-prostaglandin F1 α and nitrite/nitrate in the aortic ring effluent, inhibition of platelets (blunted thromboxane B2 generation), and inhibition of systemic inflammation (lower plasma concentration of serum amyloid P, haptoglobin). NicA treatment resulted in an approximately 2-fold higher concentration of MNA and its metabolites in urine and a 4-fold higher nicotinamide/MNA ratio in plasma, compared with MNA treatment. In summary; MNA displays pronounced antiatherosclerotic action in ApoE/LDLR(-/-) mice, an effect associated with an improvement in prostacyclin- and nitric oxide-dependent endothelial function, inhibition of platelet activation, inhibition of inflammatory burden in plaques, and diminished systemic inflammation. Despite substantially higher MNA availability after NicA treatment, compared with an equivalent dose of MNA, the antiatherosclerotic effect of NicA was not stronger. We suggest that detrimental effects of NicA or its metabolites other than MNA may limit beneficial effects of NicA-derived MNA.

The mechanisms underlying nitrite-induced effects on thrombosis and hemostasis in vivo are not clear. The goal of the work described here was to investigate the role of xanthine oxidoreductase (XOR) in the anti-platelet and anti-thrombotic activities of nitrite in rats in vivo. Arterial thrombosis was induced electrically in rats with renovascular hypertension by partial ligation of the left renal artery. Sodium nitrite (NaNO2, 0.17 mmol/kg twice daily for 3 days, p.o) was administered with or without one of the XOR-inhibitors: allopurinol (ALLO) and febuxostat (FEB) (100 and 5 mg/kg, p.o., for 3 days). Nitrite treatment (0.17 mmol/kg), which was associated with a significant increase in NOHb, nitrite/nitrate plasma concentration, resulted in a substantial decrease in thrombus weight (TW) (0.48 ± 0.03 mg vs. vehicle [VEH] 0.88 ± 0.08 mg, p 2 generation was fully reversed by both XOR-inhibitors. In addition, nitrite decreased plasminogen activator inhibitor-1 concentration (0.47 ± 0.13 ng/ml vs. VEH 0.62 ± 0.04 ng/ml, p 2) and normoxia (20%O2). Nitrite treatment had no effect on coagulation parameters. In conclusion, the nitrite-induced anti-platelet effect in rats in vivo is mediated by XOR, but XOR does not fully account for the anti-thrombotic effects of nitrite.

The purpose of this study was to analyse the influence of simultaneous pancreas-kidney or kidney transplantation on endothelial function and systemic inflammation in type 1 diabetic patients with end-stage renal disease. In 39 simultaneous pancreas-kidney, 39 type 1 diabetic kidney and 52 non-diabetic kidney recipients, flow-mediated dilatation was measured. Additionally, blood glycated haemoglobin, serum creatinine and lipids, plasma nitrites [Formula: see text] and nitrates, asymmetric dimethylarginine, soluble vascular cell adhesion molecule-1, intercellular adhesion molecule-1, and E-selectin, high-sensitivity C-reactive protein, tumour necrosis factor-α, interleukin 1β and interleukin 6 concentrations were assessed. During 58 ± 31 months follow-up period, flow-mediated dilatation and [Formula: see text] were greater in simultaneous pancreas-kidney than in type 1 diabetic kidney recipients [10.4% ± 4.7% vs 7.7% ± 4.2%, p0.05 and 0.94 (0.74-1.34) vs 0.24 (0.20-0.43) μmol/L, p0.01, respectively]. In type 1 diabetic patients after simultaneous pancreas-kidney or kidney transplantation, [Formula: see text] correlated with flow-mediated dilatation (r = 0.306, p0.05) and with blood glycated haemoglobin (r = -0.570, p0.001). The difference in [Formula: see text] was linked to blood glycated haemoglobin and estimated glomerular filtration rate, whereas the difference in flow-mediated dilatation was linked to [Formula: see text]. The levels of inflammatory markers (except soluble vascular cell adhesion molecule-1) were similar in simultaneous pancreas-kidney and type 1 diabetic kidney recipients. Improved endothelial function in type 1 diabetic patients with end-stage renal disease after simultaneous pancreas-kidney compared to kidney transplantation is associated with normalisation of glucose metabolism but not with improvement in plasma pro-inflammatory cytokines.

The objective of this study was to develop and validate the method based on micro-liquid chromatography-tandem mass spectrometry (microLC/MS-MRM) for simultaneous determination of adiponectin (ADN), von Willebrand factor (vWF), soluble form of vascular cell adhesion molecule 1 (sVCAM-1), soluble form of intercellular adhesion molecule 1 (sICAM-1) and syndecan-1 (SDC-1) in mouse plasma. The calibration range was established from 2.5pmol/mL to 5000pmol/mL for ADN; 5pmol/mL to 5000pmol/mL for vWF; 0.375pmol/mL to 250pmol/mL for sVCAM-1 and sICAM-1; and 0.25pmol/mL to 250pmol/mL for SDC-1. The method was applied to measure the plasma concentration of selected proteins in mice fed high-fat diet (HFD), and revealed the pro-thrombotic status by increased concentration of vWF (1.31$\pm$0.17 nmol/mL (Control) vs 1.98$\pm$0.09 nmol/mL (HFD), p

Artykuł pokazuje znaczenie studiów podyplomowych w procesie kształtowania kompetencji menedżerskich. Studia podyplomowe posiadają już długą tradycję w polskim systemie kształcenia i doskonalenia zawodowego, ale w ostatnich latach szczególnie widoczny jest rozwój tej formy w odniesieniu do pracowników na stanowiskach kierowniczych oraz adeptów zawodu menedżera. W tym artykule określono miejsce studiów podyplomowych wśród innych form doskonalenia menedżerów oraz przedstawiono analizę opinii uczestników studiów podyplomowych prowadzonych przez Katedrę Zarządzania Politechniki Łódzkiej., Zeszyty Naukowe. Organizacja i Zarządzanie, Vol. 68 No. 1217 (2017): Zeszyty Naukowe. Organizacja i Zarządzanie

Acute inhibition of NOS by L-NAME ($N^{\omega}$-nitro-L-arginine methyl ester) is known to decrease maximal oxygen consumption ($V'O_{2max}$) and impair maximal exercise capacity, whereas the effects of chronic L-NAME treatment on $V'O_{2max}$ and exercise performance have not been studied so far. In this study, we analysed the effect of L-NAME treatment, (LN2 and LN12, respectively) on $V'O_{2max}$ and exercise capacity (in maximal incremental running and prolonged sub-maximal incremental running tests), systemic NO bioavailability (plasma nitrite ($NO_{2}^{-}$) and nitrate ($NO_{3}^{-}$)) and prostacyclin ($PGI_{2}$) production in C57BL6/J mice. Mice treated with L-NAME for 2 weeks (LN2) displayed higher $V'O_{2max}$ and better running capacity than age-matched control mice. In LN2 mice, NO bioavailability was preserved, as evidenced by maintained $NO_{2}^{-}$ plasma concentration. $PGI_{2}$ production was activated (increased 6-keto-$PGF_{1\alpha}$ plasma concentration) and the number of circulating erythrocytes (RBC) and haemoglobin concentration were increased. In mice treated with L-NAME for 12 weeks (LN12), NO bioavailability was decreased (lower $NO_{2}^{-}$ plasma concentration), and 6-keto-$PGF_{1\alpha}$ plasma concentration and RBC number were not elevated compared to age-matched control mice. However, LN12 mice still performed better during the maximal incremental running test despite having lower $V'O_{2max}$. Interestingly, the LN12 mice showed poorer running capacity during the prolonged sub-maximal incremental running test. To conclude, short-term (2 weeks) but not long-term (12 weeks) treatment with L-NAME activated robust compensatory mechanisms involving preservation of NO2- plasma concentration, overproduction of $PGI_{2}$ and increased number of RBCs, which might explain the fully preserved exercise capacity despite the inhibition of NOS.

Strenuous physical exercise leads to platelet activation that is normally counterbalanced by the production of endothelium-derived anti-platelet mediators, including prostacyclin (PGI2) and nitric oxide (NO). However, in the case of endothelial dysfunction, e.g. in atherosclerosis, there exists an increased risk for intravascular thrombosis during exercise that might be due to an impairment in endothelial anti-platelet mechanisms. In the present work, we evaluated platelet activation at rest and following a single bout of strenuous treadmill exercise in female ApoE/LDLR-/- mice with early (3-month-old) and advanced (7-month-old) atherosclerosis compared to female age-matched WT mice. In sedentary and post-exercise groups of animals, we analyzed TXB2 generation and the expression of platelet activation markers in the whole blood ex vivo assay. We also measured pre- and post-exercise plasma concentration of 6-keto-PGF1α, nitrite/nitrate, lipid profile, and blood cell count. Sedentary 3- and 7-month-old ApoE/LDLR-/- mice displayed significantly higher activation of platelets compared to age-matched wild-type (WT) mice, as evidenced by increased TXB2 production, expression of P-selectin, and activation of GPIIb/IIIa receptors, as well as increased fibrinogen and von Willebrand factor (vWf) binding. Interestingly, in ApoE/LDLR-/- but not in WT mice, strenuous exercise partially inhibited TXB2 production, the expression of activated GPIIb/IIIa receptors, and fibrinogen binding, with no effect on the P-selectin expression and vWf binding. Post-exercise down-regulation of the activated GPIIb/IIIa receptor expression and fibrinogen binding was not significantly different between 3- and 7-month-old ApoE/LDLR-/- mice; however, only 7-month-old ApoE/LDLR-/- mice showed lower TXB2 production after exercise. In female 4-6-month-old ApoE/LDLR-/- but not in WT mice, an elevated pre- and post-exercise plasma concentration of 6-keto-PGF1α was observed. In turn, the pre- and post-exercise plasma concentrations of nitrite (NO2-) and nitrate (NO3-) were decreased in ApoE/LDLR-/- as compared to that in age-matched WT mice. In conclusion, we demonstrated overactivation of platelets in ApoE/LDLR-/- as compared to WT mice. However, platelet activation in ApoE/LDLR-/- mice was not further increased by strenuous exercise, but was instead attenuated, a phenomenon not observed in WT mice. This phenomenon could be linked to compensatory up-regulation of PGI2-dependent anti-platelet mechanisms in ApoE/LDLR-/- mice.

Koopetycję konstytuują dwie istotne siły: presja konkurencji i pragnienie współpracy. Pojęciem tym określa się współdziałanie podmiotów pozostających w tym samym czasie w relacjach konkurencyjnych. Presja na innowacyjność, złożoność i wysoki poziom zaawansowania technologicznego produktów oraz heterogeniczność i unikalność zasobów warunkują relacje koopetycyjne w firmach high-technology. Na podstawie badań w 402 przedsiębiorstwach high-tech stwierdzono, że posiadane zasoby i celowe tworzenie ich nadmiaru (redundancja) stanowią silną determinantę tworzenia przez nie relacji koopetycyjnych. Im więcej zasobów posiada przedsiębiorstwo, tym chętniej jednocześnie współdziała i konkuruje z rywalami, przy czym zasoby te silniej wpływają na intensywność działań kooperacyjnych niż konkurencyjnych w ramach koopetycji. (abstrakt oryginalny)

Purpose To analyze the effect of liver steatosis and obesity on pharmacokinetic profile of two structurally-related liver-selective NO-donors – V-PYRRO/NO and V-PROLI/NO. Methods C57BL/6 mice were fed control or high-fat diet for 15 weeks to induced liver steatosis and obesity (HFD mice). Pharmacokinetics and renal elimination studies were conducted in vivo following iv dosing of V-PYRRO/NO and V-PROLI/NO (0.03 mmol/kg). Hepatic clearance was evaluated ex vivo in the isolated perfused mice liver and in vitro with the use of liver microsomes. Results V-PYRRO/NO and V-PROLI/NO, despite similar structure, displayed different pharmacokinetic properties. V-PYRRO/NO was uptaken and metabolized by the liver, while V-PROLI/NO was eliminated unchanged with urine. In HFD mice, despite increased CYP450 metabolism of V-PYRRO/NO the elimination rate was slower most likely due to the impairment of hepatic microcirculation caused by liver fat accumulation. In turn, in HFD mice renal clearence of V-PROLI/NO was accelerated and volume of distribution was increased most likely due to additional intracellular water in HFD mice. Conclusions The pharmacokinetics of V-PROLI/NO, the novel proline-based analog of V-PYRRO/NO with additional single carboxylic acid moiety, attached to the molecule of V-PYRRO/NO to improve the water solubility, was differently affected by liver steatosis and obesity as compared with the parent compound V-PYRRO/NO.

Aim: Increased risk of pulmonary hypertension and not clear response of pulmonary endothelium to diabetes were reasons to compare nitric oxide (NO) and PGI-dependent function in pulmonary and systemic circulation in diabetes type II. Methods: In fasted db mice (male, 20 weeks) we compared the function of pulmonary (isolated perfused lung-IPL) and aortic endothelium (wire myograph). It was supported by determination of nitrite(NO 2 - )/nitrate(NO 3 - ), PGI production, immunostaining (IMS) and western blot. Results: In diabetic IPL NO-dependent modulation of hypoxic pulmonary vasoconstriction was impaired but filtration coefficient increased; response to U46619 was enhanced; cumulative generation of NO 2 - was decreased (NO 3 - , basals not changed) but 6-keto-PGF 1α increased. In diabetic lungs CD141 but not eNOS IMS was increased, fibrosis of parenchyma was weak and ultrastructural endothelial changes pronounced. In diabetic aortic rings impairment of acetylcholine-induced vasodilation, ionophore-stimulated NO 2 - (not basal) and 6-keto-PGF 1α production were observed. CD141 but not eNOS IMS was lower. In diabetes general protein nitration and nitration of PGI synthase (PGIS) were substantially increased in aorta but not in the lungs. Conclusions: Advanced diabetes affects pulmonary circulation resulting in (1) increased permeability of microcirculation and vasoreactivity; (2) impairment of NO-dependent vascular function paralleled with aorta; (3) increased PGI production contrasted with aorta; (4) weak nitration of pulmonary PGIS as oppose to systemic. In the diabetic lungs impairment of NO-dependent function was not alongside production of PGI; it may be due to the relative resistance to non-enzymatic nitration of PGIS.

Background: Pulmonary arterial hypertension (PAH) is associated with inflammatory response but it is unknown whether it is associated with alterations in NNMT activity and MNA plasma concentration. Here we examined changes in NNMT-MNA pathway in PAH in rats and humans. Methods: PAH in rats was induced by a single subcutaneous injection of MCT (60mg/kg). Changes in NNMT activity in the lungs and liver (assessed as the rate of conversion of nicotinamide (NA) to MNA), changes in plasma concentration of MNA and its metabolites (analyzed by LC/MS) were analyzed in relation to PAH progression. PAH was characterized by right ventricular hypertrophy (gross morphology), cardiac dysfunction (by MRI), lung histopathology, lung ultrastructure, and ET-1 concentration in plasma. NO-dependent and PGI2-dependent function in isolated lungs was analyzed. In naive patients with idiopathic pulmonary hypertension (IPAH) characterized by hemodynamic and biochemical parameters MNA and its metabolites in plasma were also measured. Results: MCT-injected rats developed hypertrophy and functional impairment of the right ventricle, hypertrophy of the pulmonary arteries, endothelial ultrastructural defects and a progressive increase in ET-1 plasma concentration-findings all consistent with PAH development. In isolated lung, NO-dependent regulation of hypoxic pulmonary vasoconstriction was impaired, while PGI2 production (6-keto-PGF1α) was increased. NNMT activity increased progressively in the liver and in the lungs following MCT injection, and NNMT response was associated with an increase in MNA and 6-keto-PGF1α concentration in plasma. In IPAH patients plasma concentration of MNA was elevated as compared with healthy controls. Conclusions: Progression of pulmonary hypertension is associated with the activation of the NNMT-MNA pathway in rats and humans. Given the vasoprotective activity of exogenous MNA, which was previously ascribed to PGI2 release, the activation of the endogenous NNMT-MNA pathway may play a compensatory role in PAH.

What is the central question of this study? The main aim of the present study was to determine the effect of prolonged moderate-intensity endurance training on the endothelial glycocalyx layer integrity in relationship to the training-induced changes in oxidative stress and antioxidant defence in humans. What is the main finding and its importance? We have shown, for the first time, a protective effect of prolonged moderate-intensity endurance training on endothelial glycocalyx layer integrity, as judged by significantly lower basal and end-exercise serum concentrations of glycocalyx damage markers, i.e. syndecan-1 and heparan sulfate, accompanied by attenuation of oxidative stress and enhancement of antioxidant defence after training in previously untrained healthy young men. In this study, we evaluated the effect of 20 weeks of moderate-intensity endurance training (ET) on the endothelial glycocalyx layer integrity in relationship to the training-induced changes in antioxidant defence. Eleven healthy young, untrained men performed an incremental cycling exercise bout until exhaustion before and after 20 weeks of ET. Endurance training consisted of 40 min sessions, mainly of moderate intensity (∼50% of maximal oxygen uptake), performed four times per week. Venous blood samples were taken at rest and at the end of the maximal exercise test. Muscle biopsies from vastus lateralis were taken before and after the training. Endurance training resulted in a significant increase in physical capacity (P 0.05) as reflected by an increase in power output reached at the lactate threshold and at maximal oxygen uptake. Training led to a decrease (P 0.05) in basal and end-exercise concentrations of blood markers of glycocalyx damage (syndecan-1 and heparan sulfate). The lowering of glycocalyx shedding after the ET was accompanied by an attenuation of oxidative stress, as evidenced by a decrease in the basal plasma concentration of isoprostanes, and by an increase in antioxidant defence, reflected by an enhancement in superoxide dismutase 2 protein content in vastus lateralis (P 0.05). In contrast, training did not induce a significant increase in basal nitrite/nitrate plasma concentration (P 0.05). Moderate-intensity ET exerts a pronounced protective effect on endothelial glycocalyx integrity at rest and during exercise, probably through an improvement of antioxidant defence that may represent the vasoprotective mechanisms highly responsive to moderate-intensity endurance training.

A sensitive and specific liquid chromatography electrospray ionization–mass spectrometry method for determination of 1,4-dimethylpyridinium (1,4-DMP) in rat plasma has been developed and validated. Chromatography was performed on an Aquasil C18 analytical column (4.6 × 150 mm, 5 µm, Thermo Scientific, Rockford, IL, USA) with isocratic elution using a mobile phase containing acetonitrile and water with an addition of 0.1% of formic acid. Detection was achieved by an Applied Biosystems MDS Sciex (Concord, Ontario, Canada) API 2000 triple quadrupole mass spectrometer. Electrospray ionization was used for ion production. The limit of detection in the single ion monitoring mode was found to be 10 ng/mL. The limit of quantification was 50 ng/mL. The precision and accuracy for both within-day and between-day determination of 1,4-dimethylpyridinium was 2.4–7.56 and 90.93–111.48%. The results of this analytical method validation allow pharmacokinetic studies to be carried out in rats. The method was used for the pilot study of the pharmacokinetic behavior of 1,4-DMP in rats after intravenous administration. Copyright © 2012 John Wiley & Sons, Ltd.

The efficiency of carbon adsorbents in the extraction of impurities from 1‐phenylethylamine (a model substance) and MDMA (3,4‐methylenedioxymethamphetamine), the main psychoactive component of ‘ecstasy’ tablets, was investigated. MDMA was synthesized according to two different ways in which piperonal and isosafrole were used as precursors. Three types of carbon absorbents were tested: Envicarb (surf. area 98 m2/g, Supelco), Carboprep (surf. area 400 m2/g, Restek Corporation), and Hypercarb (surf. area 94 m2/g, Thermo Electron Corporation UK). The efficiency of extraction was studied by thin layer chromatography (TLC) with UV detection (254 and 366 nm). The separation of impurities was carried out on silica gel plates with fluorescent indicator F254 and the mixture of chloroform:methanol:acetonitrile (5∶2∶3 v/v/v) was used as TLC eluent. The elaborated profiling procedure enables distinction between samples of MDMA obtained according to different synthesis methods.

Aims Extracellular nucleotides and adenosine that are formed or degraded by membrane-bound ecto-enzymes could affect atherosclerosis by regulating the inflammation and thrombosis. This study aimed to evaluate a relation between ecto-enzymes that convert extracellular adenosine triphosphate to adenine dinucleotide phosphate, adenosine monophosphate, adenosine, and inosine on the surface of the vessel wall with the severity or progression of experimental and clinical atherosclerosis. Furthermore, we tested whether the inhibition of adenosine deaminase will block the development of experimental atherosclerosis. Methods and results Vascular activities of ecto-nucleoside triphosphate diphosphohydrolase 1, ecto-5’-nucleotidase, and ecto-adenosine deaminase (eADA) were measured in aortas of apolipoprotein E-/- low density lipoprotein receptor (ApoE-/-LDLR-/-) and wild-type mice as well as in human aortas. Plaques were analysed in the entire aorta, aortic root, and brachiocephalic artery by Oil-Red O and Orcein Martius Scarlet Blue staining and vascular accumulation of macrophages. The cellular location of ecto-enzymes was analysed by immunofluorescence. The effect of eADA inhibition on atherosclerosis progression was studied by a 2-month deoxycoformycin treatment of ApoE-/-LDLR-/- mice. The vascular eADA activity prominently increased in ApoE-/-LDLR-/- mice when compared with wild type already at the age of 1 month and progressed along atherosclerosis development, reaching a 10-fold difference at 10 months. The activity of eADA correlated with atherosclerotic changes in human aortas. High abundance of eADA in atherosclerotic vessels originated from activated endothelial cells and macrophages. There were no changes in ecto-nucleoside triphosphate diphosphohydrolase 1 activity, whereas ecto-5’-nucleotidase was moderately decreased in ApoE-/-LDLR-/- mice. Deoxycoformycin treatment attenuated plaque development in aortic root and brachiocephalic artery of ApoE-/-LDLR-/- mice, suppressed vascular inflammation and improved endothelial function. Conclusions This study highlights the importance of extracellular nucleotides and adenosine metabolism in the atherosclerotic vessel in both experimental and clinical setting. The increased eADA activity marks an early stage of atherosclerosis, contributes to its progression and could represent a novel target for therapy.

Carbon monoxide (CO) modulates mitochondrial respiration, but the mechanisms involved are not completely understood. The aim of the present study was to investigate the acute effects of CO on bioenergetics and metabolism in intact EA.hy926 endothelial cells using live cell imaging techniques. Our findings indicate that CORM-401, a compound that liberates CO, reduces ATP production from glycolysis, and induces a mild mitochondrial depolarization. In addition, CO from CORM-401 increases mitochondrial calcium and activates complexes I and II. The subsequent increase in mitochondrial respiration leads to ATP production through oxidative phosphorylation. Thus, our results show that nonactivated endothelial cells rely primarily on glycolysis, but in the presence of CO, mitochondrial Ca2+ increases and activates respiration that shifts the metabolism of endothelial cells from glycolysis- to oxidative phosphorylation-dependent ATP production.

In the present study we aimed to evaluate whether oxidative stress and inflammation induced by strenuous exercise affect glycocalyx integrity and endothelial function. Twenty one young, untrained healthy men performed a maximal incremental cycling exercise – until exhaustion. Markers of glycocalyx shedding (syndecan-1, heparan sulfate and hyaluronic acid), endothelial status (nitric oxide and prostacyclin metabolites – nitrate, nitrite, 6-keto-prostaglandin F1α), oxidative stress (8-oxo-2’-deoxyguanosine) and antioxidant capacity (uric acid, non-enzymatic antioxidant capacity) as well as markers of inflammation (sVCAM-1 and sICAM-1) were analyzed in venous blood samples taken at rest and at the end of exercise. The applied strenuous exercise caused a 5-fold increase in plasma lactate and hypoxanthine concentrations (p

Endothelial dysfunction has prognostic, diagnostic and therapeutic significance in cardiovascular disease, but the endothelial phenotype is still not measured routinely to stratify the cardiovascular risk and tailor therapy. Flow-mediated dilation (FMD), the gold-standard technique for the functional assessment of endothelial function that is increasingly used in clinical settings measures the nitric oxide (NO)-dependent function only. However, the endothelial dysfunction involves a plethora of pathophysiologically important biochemical changes beyond alterations in the NO bioavailability. Still, in many diseases, some plasma protein biomarkers reflecting the pro-thrombotic and the pro-inflammatory endothelial phenotypes have poor selectivity, specificity, and a weak predictive value if they are used alone. Therefore, a multi biomarker strategy seems to be a reasonable and promising alternative. Here, we propose a multi-biomarker strategy to diagnose the endothelial dysfunction and to monitor the efficacy of an endothelium-targeted therapy. This strategy is based on the panel of endothelial biomarkers, reflecting various aspects and mechanisms of dysfunctional endothelium. The potential of an advanced analytical platform like the ultra-high performance liquid chromatography (UHPLC) coupled to mass spectrometry-based multiple reaction monitoring for simultaneous quantification of multiple endothelial biomakers is also discussed.

V-PYRRO/NO [O(2)-vinyl-1-(pyrrolidin-1-yl) diazen-1-ium-1,2-diolate] and V-PROLI/NO (O2-vinyl-[2-(carboxylato) pyrrolidin-1-yl]diazen-1-ium-1,2-diolate), two structurally similar diazeniumdiolate derivatives, were designed as liver-selective prodrugs that are metabolized by cytochrome P450 isoenzymes, with subsequent release of nitric oxide (NO). Yet, their efficacy in the treatment of nonalcoholic fatty liver disease (NAFLD) and their comparative pharmacokinetic and metabolic profiles have not been characterized. The aim of the present work was to compare the effects of V-PYRRO/NO and V-PROLI/NO on liver steatosis, glucose tolerance, and liver fatty acid composition in C57BL/6J mice fed a high-fat diet, as well as to comprehensively characterize the ADME (absorption, distribution, metabolism and excretion) profiles of both NO donors. Despite their similar structure, V-PYRRO/NO and V-PROLI/NO showed differences in pharmacological efficacy in the murine model of NAFLD. V-PYRRO/NO, but not V-PROLI/NO, attenuated liver steatosis, improved glucose tolerance, and favorably modified fatty acid composition in the liver. Both compounds were characterized by rapid absorption following i.p. administration, rapid elimination from the body, and incomplete bioavailability. However, V-PYRRO/NO was eliminated mainly by the liver, whereas V-PROLI/NO was excreted mostly in unchanged form by the kidney. V-PYRRO/NO was metabolized by CYP2E1, CYP2C9, CYP1A2, and CYP3A4, whereas V-PROLI/NO was metabolized mainly by CYP1A2. Importantly, V-PYRRO/NO was a better NO releaser in vivo and in the isolated, perfused liver than V-PROLI/NO, an effect compatible with the superior antisteatotic activity of V-PYRRO/NO. In conclusion, V-PYRRO/NO displayed a pronounced antisteatotic effect associated with liver-targeted NO release, whereas V-PROLI/NO showed low effectiveness, was not taken up by the liver, and was eliminated mostly in unchanged form by the kidney.

The study analyzed hypertension management and control rates among non-dialysis, non-transplanted hypertensive chronic kidney disease (CKD) patients under specialized care in Gdansk nephrology center in 1996-2011.It was a retrospective, cross-sectional study analyzing data from medical records of 190, 490, 1799 and 1696 subjects with CKD, who received outpatient care in 1996, 2001, 2006 and 2011, and were included in four independent surveys, respectively.The average number of antihypertensive drugs per patient increased significantly (p0.01) as follows 1.74 ± 0.9 (1996), 2.08 ± 1.01 (2011), 2.5 ± 1.19 (2006) and 2.65 ± 1.18 (2011). The percentage of patients receiving diuretics, beta-blockers and drugs inhibiting renin-angiotensin-aldosterone increased significantly in subsequent years, while a frequency of therapy with calcium channel blockers decreased (p0.001). 16%, 30%, 42% and 54% of subjects had causal BP values140/90 mmHg (p0.001). When specific thresholds for CKD patients according to JNC recommendations were used, the control rate was worse but also showed significant improvement in the second, third and final surveys, i.e. 9%, 12%, 14% and 24% (p0.001). The subgroup analysis revealed that a better control rate was observed in following groups:65 years old; I-II stage of CKD; primary glomerulonephritis; without cardiovascular complications or diabetes.The study may show an improvement in the effectiveness of antihypertensive treatment in CKD patients under specialized care in Gdansk Nephrology Centre in 1996-2011.

A sensitive and specific liquid chromatography electrospray ionization-mass spectrometry method for determination of 1,4-dimethylpyridinium (1,4-DMP) in rat plasma has been developed and validated. Chromatography was performed on an Aquasil C(18) analytical column (4.6 × 150 mm, 5 µm, Thermo Scientific, Rockford, IL, USA) with isocratic elution using a mobile phase containing acetonitrile and water with an addition of 0.1% of formic acid. Detection was achieved by an Applied Biosystems MDS Sciex (Concord, Ontario, Canada) API 2000 triple quadrupole mass spectrometer. Electrospray ionization was used for ion production. The limit of detection in the single ion monitoring mode was found to be 10 ng/mL. The limit of quantification was 50 ng/mL. The precision and accuracy for both within-day and between-day determination of 1,4-dimethylpyridinium was 2.4-7.56 and 90.93-111.48%. The results of this analytical method validation allow pharmacokinetic studies to be carried out in rats. The method was used for the pilot study of the pharmacokinetic behavior of 1,4-DMP in rats after intravenous administration.

Plasma protein binding of drugs may have significant effect on its pharmacodynamic, toxicological and pharmacokinetic properties, since only the free drug can pass across biological membrane and get to its specific site of action. Many drugs show a high affinity to albumin which is the most abundant plasma protein. In the present study capillary electrophoresis in the frontal analysis mode (CE/FA), as promising technique for assessment of drug-protein interaction was used. The free drug concentration was measured from height of the frontal peak and calculated based on the external drug standard in absence of protein. With a known concentration of total drug, the percentage of protein bound drug was determined. The binding parameters were also estimated based on the equilibrium dialysis experiment which is considered to be a reference method. This study was designed to examine the interaction of dexamethasone sodium phosphate (DXM) with BSA and HSA under simulated physiological conditions (pH 7.4, 67 mM phosphate buffer, I = 0.17). Using fixed, at physiological level, HSA and BSA concentrations and increasing DXM concentrations, the number of binding sites (n) and binding constant (K(a) ) was calculated from both nonlinear regression fitting and Scatchard Plot. Despite some differences, it can be concluded that the CE/FA is comparable with equilibrium dialysis, but since the first one offers advantages such as low sample consumption, short analysis time, and high separation efficiency, it can be used in high-throughput screening of drug protein binding at the early stage of drug discovery. Interspecies differences in binding of a drug to albumins have been observed and it should be taken into account in interpretation of the results.

The aim of the study was the estimation of the frequency intake and energy value of breakfasts and lunches consumed by school children. The group of 463 pupils of elementary and grammar schools selected at random from Pila was investigated in may 2008. Most of investigated pupils had declared the everyday consumption of breakfasts, however only half of them consumed it at home before turned out to school. Also lunch was consumed by the most of investigated children. Frequency breakfast intake decreased at older schoolchildren, but at a time the percentage of pupils preparing it individually increased, what suggests the diminution of the care of parents over the adolescent child. The energy value of consumed breakfasts and lunches was irregular Breakfasts characterized with too low, and lunches too high, participation of the energy compared to recommended daily dietary allowances. Consumed breakfasts were also incorrectly composed Breakfast vegetable and fruits and milk and its products consumption was too low.

A variety of approaches are employed for treatment of Chiari I malformation. They differ in extent of cranio-c ervical decompression. A technique based on arachnoid preservation and duroplasty was introduced in our department in 2001. The aim of the study is to compare between the previous and the present technique.Retrospective analysis of 38 patients with Chiari I malformation treated between 1998 and 2004 was performed. The previous technique including arach- noid incision, coagulation of cerebellar tonsils, and fourth ventricle exploration without duroplasty was used to treat 21 patients (group 1). A further 17 patients were treated with the present technique consisting of arachnoid preservation and duroplasty (group 2). Complication rates as well as early and late results of treatment were evaluated. Karnofsky (KPS), Rankin (RS) and Bidziński (BS) scales were used for evaluation of results.Post-operative complications were detected in 9 patients in group 1 (43%). They included liquorrhoea (5 cases), meningitis (1 case), and symptoms progression (3 cases). There were no surgical complications in group 2 (p = 0.002). Neurological improvement in the early period (until discharge from hospital) occurred in 10 (48%) patients in group 1 and in 13 (76%) in group 2 (p = NS). Further improvement or lack of symptoms progression was found in 58% in group 1 and 82% in group 2 (p = NS). Assessment in KPS, RS and BS showed slightly better results of group 2 but the difference was statistically insignificant.Results of both techniques are comparable. The risk of post-operative complications after extra-arachnoid cranio-cervical decompression with duroplasty is, however, significantly lower.

A variety of methods have been employed for syrinx drainage, especially for treatment of syringomyelia unrelated to Chiari 1 malformation (C1M). Research on CSF circulation revealed that perpetual syrinx increase may result from disturbances of CSF flow and transient hypertension within the spinal subarachnoid space. Thus the concept of CSF drainage out of the spinal canal has arisen. The aim of the study is to present the outcome of patients with syringomyelia treated by placement of a syringoperitoneal shunt (SPS).An SPS was implanted in 5 patients with syringomyelia between 2004 and 2007. The syrinx was secondary to C1M in 2 patients, post-inflammatory in another 2 and idiopathic in 1 case. Myelotomy was performed over the widest part of the syrinx, at the lowest possible level. The proximal end of the drain was introduced into the syrinx rostrally, and the distal end to the peritoneum. For outcome evaluation late clinical and radiological results of treatment were used. The follow-up ranged from 1 to 4 years.Clinical improvement or stabilization of symptoms was observed in 4 patients. One patient was re-operated on due to the drain slipping out of the syrinx. Despite syrinx decrease on follow-up MR scan, further clinical deterioration led to re-exploration of the cranio-cervical junction. Follow-up MR scans revealed syrinx decrease in all but 1 patient with C1M.Preliminary results suggest that SPS is a safe and efficient treatment method for patients with syringomyelia not related to structural anomalies at the cranio-vertebral junction. For treatment of syringomyelia secondary to C1M following failure of cranio-cervical decompression, another exploration of this area should be considered.

The aim of the study is to present the authors' experience with the treatment of metastatic tumours of the thoracic spine regarding surgical strategy depending on extent of neoplastic invasion.Between January 2002 and August 2005, 15 patients with thoracic spinal tumours underwent surgical treatment with instrumental stabilization at the Department of Neurosurgery, Warsaw Medical School, Poland. Seven patients with tumours localized in vertebral bodies or vertebral bodies and pedicles were operated on via an anterior approach with concomitant stabilization at the same operative procedure. In 5 patients with metastatic involvement of vertebral arches and pedicles, a posterior approach for tumour removal was used with concomitant posterior fixation at the same operative procedure. Two patients with multiple spine metastases underwent internal fixation at the level corresponding to the observed symptoms. In 1 patient with bilateral lung cancer and vertebral body involvement, posterior stabilization with decompression of nervous structures was performed.None of the patients neurologically deteriorated after surgery. All patients with neurological deficits improved while one patient with pain syndrome did not. In 1 case approach-related surgical complications were observed.Regarding the complexity of surgical treatment of spinal metastases, the presented material is clearly not sufficient to draw firm conclusions. However, according to the authors' experience, the treatment of choice in single spinal metastasis involving the vertebral body is tumour removal via an anterior approach followed by adjunctive therapy. In metastatic lesions localized in vertebral pedicles and arches, removal via a posterior approach with concomitant posterior stabilization at the same operative procedure and adjunctive therapy is most indicated. Posterior stabilization of the spinal cord segment corresponding with symptoms is considered an appropriate surgical treatment of multiple spinal metastases.

Biochemical abnormalities are thought to be important factors causing bipolar disorder. Several lines of evidence suggest dopamine may play a role in its etiology, e.g. depressive syndrome is frequently encountered in subjects affected by Parkinson's disease, where dopamine depletion is observed. Thus the genes of the dopaminergic system are good candidates for studies on bipolar disorder. In the present study, we investigated polymorphisms of dopamine receptors, type D2, including the dopamine receptors D2, D3 and D4.Polish bipolar patients (n=339) and control subjects (n=366) were analyzed for three dopamine receptor gene polymorphisms: dopamine receptor D2 (DRD2) 141 ins/del, dopamine receptor D3 (DRD3) Ser9Gly, and dopamine receptor D4 (DRD4) 521 C/T.Allele and genotype frequencies did not differ significantly among patients and controls. We did not obtain any significant association between the studied polymorphisms and bipolar disorder.The results suggests that the studied gene variants of type D2 dopamine receptors are not promising candidate genes for bipolar affective illness. We did not consider the family history of the examined subjects, and also the control group was not psychiatrically screened, which may contribute to the lack of significant results.

We present the case of 81-years old woman with diabetes mellitus and moderate hypertension who reported multiple syncope's. Cerebral and metabolic causes of syncope's were excluded and the patient was diagnosed to have sick sinus syndrome. After implantation of the cardiac pacemaker the treatment with sotalol was started. After introducing the antiarrhythmic drug the frequency of syncopes increased dramatically and blood pressure rose unusually (up to 250/140 mm Hg). Subsequently phaeochromocytoma was suspected. Laboratory data as well as computed tomography confirmed the diagnosis of left adrenal tumour. After successful surgery the blood pressure normalized and the patient recovered uneventfully. The reported cases describes the difficulties in diagnosing phaeochromocytoma in elderly people with multiple different accompanying diseases.

The paper aims to investigate the relationship between strategy and structure in the high-technology enterprises. The study attempts to ascertain how chief executive officers perceive the impact of strategy on organizational structure, and likewise impact of structure on strategy, at two phases in the innovation process: the phase of innovation exploration; and the phase of innovation exploitation. The research was conducted in 61 high-technology companies based in Poland that operate either in Poland or in the global marketplace. The results show that, during the exploration of innovation, chief executive officers consider that the impact of organizational structure on strategy is stronger than the impact of strategy on structure. During the exploitation of innovations, the impact of strategy on structure is stronger.

It has been repeatedly shown that regular aerobic exercise exerts beneficial effects on incidence and progression of cancer. However, the data regarding effects of exercise on metastatic dissemination remain conflicting. Therefore, in the present study the possible preventive effects of voluntary wheel running on primary tumor growth and metastases formation in the model of spontaneous pulmonary metastasis were analyzed after orthotopic injection of 4T1 breast cancer cells into mammary fat pads of female Balb/C mice. This study identified that in the mice injected with 4T1 breast cancer cells and running on the wheels (4T1 ex) the volume and size of the primary tumor were not affected, but the number of secondary nodules formed in the lungs was significantly increased compared to their sedentary counterparts (4T1 sed). This effect was associated with decreased NO production in the isolated aorta of exercising mice (4T1 ex), suggesting deterioration of endothelial function that was associated with lower platelet count without their overactivation. This was evidenced by comparable selectin P, active GPIIb/IIIa expression, fibrinogen and vWF binding on the platelet surface. In conclusion, voluntary wheel running appeared to impair, rather than improve endothelial function, and to promote, but not decrease metastasis in the murine orthotopic model of metastatic breast cancer. These results call for revising the notion of the persistent beneficial effects of voluntary exercise on breast cancer progression, though further studies are needed to elucidate mechanisms involved in pro-metastatic effects of voluntary exercise.