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The influence of LDL-apheresis on aggregation of blood platelets, blood coagulation, and the effect of standard drugs.
Černotová, Veronika ; Mladěnka, Přemysl (advisor) ; Čečková, Martina (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Veronika Černotová Supervisor: prof. Přemysl Mladěnka, PharmD., Ph.D. Consultant: Lukáš Konečný, MSc. Title of diploma thesis: The influence of LDL-apheresis on aggregation of blood platelets, blood coagulation and the effect of standard drugs LDL-apheresis is a method that removes LDL-cholesterol (LDL-C) from the blood. It is used to treat familial hypercholesterolemia (FH), a genetic disorder causing high LDL-C levels and an early development of cardiovascular diseases. Blood platelets and coagulation system play an important role in these diseases and their activity is also affected by lipids. The aim of this thesis was to analyze possible differences in platelet aggregation and blood coagulation in patients suffering from FH. Two methods of treatment in this group were compared - lipid apheresis and PCSK9Ab (proprotein convertase subtilisin/kexin type 9 monoclonal antibodies). The observed parameters were also compared with age-matched healthy volunteers. Our cohort consisted of 15 patients and 15 healthy donors. Six patients were treated with lipid apheresis and also PCSK9Ab, six subjects only with PCSK9Ab. Platelet aggregation was measured with an impedance aggregometer using 7 different...
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The influence of LDL-apheresis on aggregation of blood platelets, blood coagulation, and the effect of standard drugs.
Černotová, Veronika ; Mladěnka, Přemysl (advisor) ; Čečková, Martina (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Veronika Černotová Supervisor: prof. Přemysl Mladěnka, PharmD., Ph.D. Consultant: Lukáš Konečný, MSc. Title of diploma thesis: The influence of LDL-apheresis on aggregation of blood platelets, blood coagulation and the effect of standard drugs LDL-apheresis is a method that removes LDL-cholesterol (LDL-C) from the blood. It is used to treat familial hypercholesterolemia (FH), a genetic disorder causing high LDL-C levels and an early development of cardiovascular diseases. Blood platelets and coagulation system play an important role in these diseases and their activity is also affected by lipids. The aim of this thesis was to analyze possible differences in platelet aggregation and blood coagulation in patients suffering from FH. Two methods of treatment in this group were compared - lipid apheresis and PCSK9Ab (proprotein convertase subtilisin/kexin type 9 monoclonal antibodies). The observed parameters were also compared with age-matched healthy volunteers. Our cohort consisted of 15 patients and 15 healthy donors. Six patients were treated with lipid apheresis and also PCSK9Ab, six subjects only with PCSK9Ab. Platelet aggregation was measured with an impedance aggregometer using 7 different...
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Theory and practice of using heparin flush
BAŠTÁŘOVÁ, Barbora
The bachelor thesis deals with recognition of theory and praxis of using the heparine flush in hospital establishments. The heparine, is currently registered as a risk medicament and more administration was added to it, which means that it is not used in praxis so much. These measures have been applied since after 2006, after the case which shocked the Czech Republic. Blood, blood clotting, heparine, heparine flush and venous ports in which it is or was used, are described in the theoretical part. The research part was implemented by quantitative method in the form of a survey of own creation, which was replied by 200 respondents. I chose 4 following departments for the research: internal department, department of surgery, department of oncology and department of paediatrics. The aim was to find out what is the standard of using the heparine flush in praxis. Following hypothesis were determined. Hypothesis n. 1, the frequency of using the heparine flush within 24 hours depends more on the doctor's office than on routine administering. Hypothesis n. 2, the way of solution dilution for the heparine flush depends rather on the doctor's office than on the department routine. Hypothesis n. 3, the way of administering the heparine flush by a nurse depends on nurse's experience with complication occurance. A non-standard survey was prepared to achieve the aim and to test the hypothesis. The survey was evaluated by means of graphical and statistical methods. The survey contained 25 questions. It was possible to state from the whole research, that the heparine flush has not been used so much and it is rather a risk than a benefit for a patient. An interesting information, which came up with the evaluation, was a fact, that nurses encountered the term heparine flush for the first time only at some of the departments where they begin to work. The outcomes can serve as materials for education of students at medical schools and they can improve the knowledge of the heparine flush, although it is not used so much.
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Determination of Heparin by Sequential Injectin Analysis with Spectrophotometric Detection
Lišková, Růžena ; Hraníček, Jakub (advisor) ; Nesměrák, Karel (referee)
In this work the determination of heparin using Sequential Injection Analysis (SIA) technique with spectrophotometric detection was studied. The principle of this method was measuring the decrease of absorbance of phenothiazine dyes in its maximum wavelength caused by its interaction with heparin. The decrease is proportional to concentration of heparin in sample. The aim of this work was to find optimum conditions for determination of heparin by the SIA with spectrophotometric detection and to try to apply this measurement to real sample under these conditions. Three phenothiazine dyes were used: Methylene Blue, Azure A and Azure B and for each of them an optimum concentration was found. Their values were for methylene blue 4.0 ∙ 10-5 mol/dm3 and 5.0 ∙ 10-5 mol/dm3 for Azure A and Azure B. 150 µl was chosen as an optimum amount of heparin and reagent aspirated. Optimum reaction time was 0 s and flow rate during measurement 1.5 ml/min. Working concentration range was found between 0.23 and 15 mg/dm3 for methylene blue, 1.2 and 13 mg/dm3 for Azure A and 0.67 and 12 mg/dm3 for Azure B. Limit of detection varied between 0.09 - 0.35 mg/dm3 and limit of quantification between 0.30 and 1.2 mg/dm3 for different dyes. Relative standard deviation was determined as a measure of repeatability. Its values were...
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Flow injection analysis of selected glycosaminoglycans with spectrofluorimetric detection
Tichá, Renata ; Nesměrák, Karel (advisor) ; Rychlovský, Petr (referee)
The thesis is focused on a determination of heparin and chondroitin sulfate, using flow injection analysis with spectrofluorimetric detection. The determination is based on the interaction of negatively charged heparin, chondroitin sulfate resp., with a cationic dye (azure B or phenosafranine) which is manifested by the decrease in fluorescence intensity of the dye in its emission maximum. The optimal conditions for the determination in static mode were found, and calibration dependencies were measured. The conditions of FIA were optimized and following parameters were established: the volume of dispensed sample of 100 ml, the length of the reaction coil 60 cm, the flow rate 0.7 ml min-1 , the concentration of azure B 1.6×10-5 mol dm-3 , the concentration of phenosafranine 3.5×10-5 mol dm-3 . For the determination of heparin using azure B it was found: LOD = 0.023 IU ml-1 , LOQ = 0.186 IU ml-1 , and linear dynamic range 0.19-1.43 IU ml-1 . For the determination of heparin using phenosafranine it was found: LOD = 0.102 IU ml-1 , LOQ = 0.192 IU ml-1 , and linear dynamic range 0.19-1.79 IU ml-1 . For the determination of chondroitin sulfate using azure B it was found: LOD = 0.58 mg dm-3 , LOQ = 2.37 mg dm-3 , and linear dynamic range 2.37-8.32 mg dm-3 . The developed determination was applied to the...
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The use of thromboelastography (TEG) in the evaluation of coagulation in patients on intensive care unit (ICU)
Durila, Miroslav ; Cvachovec, Karel (advisor) ; Stříteský, Martin (referee) ; Kovářová Kudrnová, Zuzana (referee)
Patients in the intensive care unit are in critical condition which is often accompanied by a coagulation disorder. Sepsis as a leading cause of death in critically ill patients may be associated with both hypercoagulable state with microtrombi formation in microcirculation and with increased production of endogenous heparinoids with inhibitory effects on blood clotting. Central venous catheter and arterial catheter are established in patients for hemodynamic monitoring and these are flushed with heparin to prevent their closure. Both inputs are used for blood sampling for laboratory tests such as blood count and coagulation parameters, including thromboelastography (TEG). In the first step of the work, arterio-venous differences in coagulation parameters were investigated in patients with sepsis. Higher concentration of D-dimers and lower antithrombin activity were found in venous blood. This finding can be explained by increased antithrombin consumption in hypercoagulable state and reactive hyperfibrinolysis. Inconsistency in the site of blood sampling may then lead to misinterpretation of the pathophysiological processes in the body. No significant differences were found in TEG parameters. In the second step of the work we examined how heparin commonly used for catheter flushing affects TEG-assessed...
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Účinnost a bezpečnost léčby NOAC
SVOBODOVÁ, Hana
Anticoagulation treatment has dramatically changed in recent years with the arrival of new molecules with different properties compared to a commonly used drugs. Massive expansion of these drugs is prevented by their high price and an indication of health insurance companies. In the first part of this thesis is described the process of blood clotting and its pathology, commonly used drugs such as heparin and warfarin, and new anticoagulants NOAC, their agents, properties, indications and contraindications.
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Characterization of protamine influence on indirect electrophoretic determination of heparin
Petrák, Ondřej ; Křížek, Tomáš (advisor) ; Kubíčková, Anna (referee)
Cílem této bakalářské práce bylo studium interakcí heparinu a protaminu v prostředí kapilární zónové elektroforézy pomocí jejich vlivu na velikost plochy píku tetraargininu. Byla použita křemenná kapilára o vnitřním průměru 50 µm a celkové délce 50 cm. Efektivní délka kapiláry byla 41,5 cm a vzorek byl dávkován na dlouhý konec kapiláry. Byl optimalizován způsob dávkování jednotlivých zón, vhodné složení základního elektrolytu bylo již známo z předešlých experimentů, na které tato práce navazovala. Základní elektrolyt byl složen z 0,1 M roztoku kyseliny trihydrogenfosforečné a 0,1% roztoku hydroxyethylcelulózy. Vzorek byl dávkován hydrodynamicky tlakem 5,0 kPa po dobu 120 s. Následně byla stejným tlakem po dobu 90 s dávkována zóna základního elektrolytu. Poslední dávkovanou zónou byla zóna tetraargininu o koncentraci 1 mg/ml, která byla dávkována hydrodynamicky tlakem 5,0 kPa po dobu 3 s. Posledním krokem bylo vložení separačního napětí 30 kV po dobu 30 s. Tetraarginin byl detegován při λ = 200 nm. Byly změřeny vlivy samotného heparinu/ protaminu a jejich směsí v různých vzájemných poměrech. Bylo ověřeno, že se zvyšující se koncentrací heparinu dochází k poklesu plochy píku tetraargininu. Nově bylo zjištěno, jaký vliv na velikost tohoto účinku má přítomnost protaminu. Zde došlo k překvapivému...
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Comparison of the efficacy of arterial flushing sets: heparinized flush versus saline flush
Strychová, Zdenka ; Di Cara, Veronika (advisor) ; Dynáková, Šárka (referee)
(v AJ) The aim of this diploma thesis was namely to find out whether the saline solution, intended for the continuous flushing of arterial catheters, is as effective in preventing arterial blockage as compared to saline solution with heparin. In this diploma thesis I also deal with the comparison of material consumption by using both methods of flushing arterial sets and the financial costs associated with them. Patients with diagnosis of sepsis or septic shock were included in the study. Pseudorandomization was used to group 52 patients into either experimental and control group. The testing was performed using a non-parametric Wilcoxon two-assay test and Fisher precision test. Based on the results of my study, regard to the occurrence of complications (catheter closure) an arterial catheter flushing by saline solution is as effective as flushing by a saline solution with heparin. My research also confirmed that the use of saline solution as a flushing solution is less costly (regarding material) and therefore it is advisable to use saline solution for flushing arterial catheters. Based on my study, saline solution is now used as a flushing solution of arterial catheters at an anesthesiology-resuscitation department, where only a saline solution with heparin was used for the flushing. Using saline...
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Indirect determination of heparin by capillary electrophoresis
Filounová, Barbora ; Křížek, Tomáš (advisor) ; Kubíčková, Anna (referee)
Heparin is a mixture of sulfonated polysaccharides which is negatively charged. Heparin is a substance which is important in organism and fundamentally affects its physiology. Main attribute of heparin is anticoagulation - it prevents the complete blood coagulation. This anticoagulant effect balances the hemocoagulation by influencing the coagulation pathway. In some cases a pharmacological application of heparin is needed so the heparin is administrated as a injection of physiological solution of sodium or calcium heparine salt. Monitoring of level of the heparin in blood is problematic - methods used today are based on the measurement of a time required for blood clot formation. The result evaluation is done by comparing a sample with reference solution. These methods are relatively imprecise, can not be used in "on-line" setting and are highly influenced by general health condition of patient. In this work some principles of affinity capillary electrophoresis were adapted from another work - heparin was determined indirectly by monitoring of decrease of the peak area of protamine. Protamine is medically used antidote of heparin because they create a stable complex together. In this work protamine was replaced by well defined tetraarginine because the most frequent amino acid in protamine is...
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