Abstract Two reversed-phase high performance liquid chromatography analytical methods (Method I and Method II) for determination of assay of recombinant human thrombin in pharmaceutical formulations were developed and validated. Analysis was performed on chromatographic system Agilent 1200 series SL with diode array detection and mass selective detection. Method I was intended for faster determination of thrombin assay. Gradient programme was optimised to achieve sufficient separation and acceptable runtime. Chromatographic analysis was performed on analytical column Grace Vydac, C4 250 × 4.6 mm, 5 mm. Method II is Method I adapted to use the mass selective detector. Chromatographic separation was performed on analytical column Zorbax 300SB-C8 SolvSaver Plus, 150 × 3 mm, 3.5 mm. Both analytical methods were validated with respect to specificity, linearity, precision and accuracy. The response of thrombin was a linear function of concentration over the range 0.1-1.0 mg/ml. Precision and accuracy of thrombin was evaluated at three concentration levels low (0.2 mg/ml), medium (0.4 mg/ml) and high (0.8 mg/ml). Both validated methods have been successfully applied for determination of assay and thrombin degradation products in pharmaceutical formulations.

Abstract This study deals with effect of hospital pharmacist on solving drug-related problems (DRP) in inpatients. The study was carried out as a prospective 5-week study at the Cardiology Clinic, Teaching Hospital, Nitra. The study group included 73 inpatients. Pharmacotherapy of each patient was analysed for DRP within 24-48 h after admission. Information on patients was collected from electronic database, medical reports, communication with attending doctors and ward rounds. Patients’ age, medical history, diagnoses, medication prescribed during hospitalisation and laboratory test results were assessed for further pharmacotherapy rationalisation. Pharmaceutical Care Network Europe (PCNE) classification scheme for DRP V6.2 was used to characterise DRP. There were 36 DRP identified in 37% inpatients (n = 27). The most frequent causes of DRP were dose-related (n = 13; 26%) and use

Abstract Among the basic rights and freedoms, stipulated in the Constitution of the Slovak Republic, are right to protection of health, free medical care based on health insurance, right to medical aids under conditions stipulated by law. These rights are guaranteed through provisions of Art. 40 of the Constitution of the Slovak Republic as part of Economic, Social and Cultural Rights and they apply to every individual. Since these rights are considered to be human rights under the international law, they are guaranteed by multilateral treaties in which Slovak Republic is a signatory, such as International Covenant on Economic, Social and Cultural Rights. Legal regulation of operating pharmacies in the Slovak Republic covers a broad scope of issues and is quite extensive. Through concerted legislative effort, the legislator had tried to set rules that would make the legal environment in this sector more transparent and easier to navigate through. In some areas, the rules were set by the legislature of the European Union (EU) and the Slovak legislature only adapted them. The most important aspect of legal regulation in this sphere that is not completely regulated by the EU legislature is providing pharmaceutical care and usage of the public health insurance system with regard to providing pharmaceutical care. The main aim of this study is to point out select issues that are associated with the position of a pharmacy operator and obligations stipulated in the Medicines Act 2011 that must be observed and incorporated into the running of a pharmacy. This area of business is fairly strictly regulated and therefore it is necessary to get acquainted with the relevant legislation. The study addresses several issues regulated by the Medicines Act 2011 or the Appropriate Pharmaceutical Practice Regulation 2012 from the legal-theoretical perspective.

Abstract Medical devices (MD), together with pharmacotherapy are supportive treatment of many acute and chronic diseases. MD as a part of medical technologies lead to more effective treatment, faster patient recovery and a reduced risk of complications. Many MD are reimbursed from public health insurance funds entirely; for others, particularly advanced functional types of MD, there must be patient participation on price or they can buy them according own decision (direct sale). The target of this study is to analyze the data from paid databases of National Center for Health Information (NCHI) that collects the outputs of provided healthcare. The most recent data were from 1.1.2013 to 30.9.2013. According to NCHI, apart from community pharmacies, there are 226 registered establishments that sold MD until 30.9.2013. Their specialization included dispensing MD (n=163), dispensing orthopaedic devices (n=48) and dispensing audioprosthetic devices (n=15). In the observed period, average monthly spending on MD were 10.1 million packages and 14 million €. Average monthly spending on reimbursed MD were 8.8 million packages and 12.9 million €. The groups with the largest shares were MD for incontinence and urinary retention, 7.7 million packages (86.9%) and 3.9 million € (30.7%); plasters and bandaging materials, 0.5 million packages (6.0%) and 1.3 million € (10.1%); MD for ostomies, 0.4 million packages (4.5%) and 1 million € (8.0%); and the MD for diabetics group, 0.1 million packages (1.4%) and 1.6 million € (12.8%). Direct sales of MD per month on average reached 1.3 million packages and 1.1 million €. The groups with largest shares were MD for incontinence and urinary retention, 629,660 packages (50.3%) and 291,919 € (26.2%); plasters and bandaging materials, 388,111 packages (31.0%) and 227,119 € (20.4%); and MD for diabetics, 56,014 packages (4.5%) and 81,721 € (7.3%).

Abstract Alkaptonuria is a rare inherited genetic disorder of phenylalanine and tyrosine metabolism. This is an autosomal recessive condition that is caused due to a defect in the enzyme homogentisate 1,2-dioxygenase, which participates in the degradation of tyrosine. As a result, homogentisic acid and its oxide accumulate in the blood and are excreted in urine in large amounts. The polymer of homogentisic acid called alkapton impregnates bradotrophic tissues.

Abstract The existing ideas about the ketorolac mechanism of analgetic potency are associated with its effect on COX isoenzymes and its significant potency needs to be explained more completely. Its impact on vanilloid receptors 1 (TRPV1) can also be an effective mechanism of analgetic action. We have evaluated the analgetic potential of ketorolac on the basis of its influence on TRPV1. It is known that Tyr511 and Ser512 amino acids are active central components of TRPV1. Thereby the synthesis of the dipeptide Tyr-Ser has been conducted as a model of the TRPV1 active centre. In our model test, using spectrofluorometry, the formation of intermolecular complexes was shown: “ketorolac - Tyr-Ser” and “capsaicin - Tyr-Ser” at stability constants Kr=0.53 • 10-4 L/mol; Kkor= 0.998 and Kr =0.3 • 10-4 L/mol; Kkor=0.998, respectively. Verification of the data was proved in experiments in vitro (the preparation of portal vein) and in vivo (Tail-flick model), with the TRPV1 agonist and antagonist. On portal vein, it was established (from initial level) by Krebs solution that the range of relaxation of smooth muscles of the portal vein (SM) was as follows at the concentration of capsaicin 0.1 μmol/L +30.3%±5.3%, capsazepine 0.5 μmol/L -3.2±2.7% and ketorolac 1.0 μmol/L -60.0±7.0%. In the experiment, at the above mentioned doses of ketorolac on the background of the action of the capsaicin (capsazepine), the range of relaxation of smooth muscles (SM) of the portal vein was -59.3±5.0% and -63.0±6.0% respectively testifying to the fact that the effect of capsaicinoids was neutralised by ketorolac. On the Tail-flick model, an atypical potentiation of the ketorolac antinociception at simultaneous use with the capsaicin was shown. The obtained data allow suggesting that TRPV1 can activate the ketorolac antinociceptive effects.

Abstract The contents of selected elements (Mn, Fe, Co, Ni, Cu, Zn, Hg and Pb) in samples of loose tea of agrimony “Repikovy čaj” (Agrimonia eupatoria L.) were studied in this work. Identification and determination of the selected elements content were performed using X-ray fluorescence spectrometry - the nuclear analytical method based on the interaction between low-energy gamma and X radiation and analysed substance. Solid form samples were pressed in tablets and directly analysed. Prepared infusions were analysed after the preconcentration of the elements using precipitation with thioacetamide and ammonium diethyldithiocarbamate with subsequent filtration of created precipitates through a nitrocellulose filter. Detection limits obtained from both methods were compared and their suitability for elemental analysis of plant preparations was discussed.

Beneficial effects of red wine polyphenols on cardiovascular health are well known. The aim of our research was an interaction study of four red wine polyphenols – resveratrol (R), quercetin (Q), kaempferol (KF) and isorhamnetin (IR) of their ability to activate endothelial NO synthase (eNOS) promoter when used alone and in equimolar mixtures. To determine their activity, we performed a luciferase reporter gene assay on EA.hy926 cells stably transfected with a luciferase reporter gene construct containing eNOS promoter. The Bradford assay was also performed to account the cytotoxicity and/or the cell number differences. The median effect equation, as an interaction analysis evaluating synergy or antagonism of the combinations was done according to mass-action law principle. Isobolographic method was performed on selected double mixtures and dose reduction index was calculated for all mixtures. All single polyphenols activated eNOS promoter. The EC50 values were in micromolar concentrations ranging from 3.44 μM (R2 = 0.96) for kaempferol to 9.89 μM for isorhamnetin (R2 = 0.94). All mixtures activated eNOS promoter, but their interactions varied from synergy (Q+R, Q+IR+KF, Q+R+KF and Q+R+IR+KF), through additive (R+IR+KF) to antagonistic interaction (R+IR, R+KF, Q+IR, Q+KF, IR+KF and R+Q+IR). In this study, we show for the first time that red wine polyphenols activated eNOS promoter when used alone and in mixtures with different type of interactions.

The micellization process of the local anaesthetic carbisocainium chloride in water was investigated by two spectral methods using the probe pyrene. First, the absorption spectroscopy in UV/VIS region was based on studying changes in characteristic absorption spectrum of pyrene in presence of surfactant. The resultant plot of the sum of absorbances for all the major pyrene peaks as a function of the total surfactant concentration shows, around the critical micellar concentration, a typical sigmoidal increase. The fluorescence emission spectroscopy in UV/VIS region of spectrum by the probe pyrene, second procedure, was applied for determination of the cmc from the measurements of the pyrene I1 /I3 ratio as a function of the surfactant concentration. The pyrene ratio data were fitted by the Boltzmann-type sigmoid of decreasing character.