|
|
Dependence of derived fibrinogen on values of DH from APTT and QUICK(PT) methods
Klus, Michal ; Zapletal,, Ondřej (referee) ; Škutková, Helena (advisor)
Hemocoagulation, blood coagulation, is an important indicator of hemostatic balance in the human body. There are many ways how to investigate blood clotting. In practice, next to the tests investigating time of coagulation cascade from view of internal way (APTT – activated partial tromboplastin time) and external way (PT – prothrombin time) is often used determining of fibrinogen concentration by Clauss method. Derived fibrinogen method determined fibrinogen concentration, too, by subtracting form the clotting curve in PT test. The reaction for Clauss method is not necessary here. Derived fibrinogen is not used much in practice. This is the reason, why the thesis related to this project will try to find relationship between concentration of fibrinogen and standard tests APTT and PT. Clinical data will be used for this.
|
|
|
Anticoagulation factors and blood uptake by monogeneans of the family Diplozoidae
Skipalová, Karolína ; Mikeš, Libor (advisor) ; Sojka, Dan (referee)
For the successful food intake by organisms that feed on blood is essentials presence of antihaemostatic molecules such as vasodilators, anticoagulant molecules and apyrases., Although members of family Diplozoidae (Heteronchoinea) are blood-feeding parasites on the gills of the fish, these molecules, that could disrupt host hemostasis, have not yet been identified. Thus, the aim of this study was to find molecules with potential anticoagulant activity in homogenates of whole worm bodies and excretory/secretory products of the members of family Diplozoidae. Furthermore perform bioinformatics analysis of sequences obtained from transcriptom project of Eudiplozoon nipponicum (Heteronchoinea: Diplozoidae) and selected proteins (protein domain) then expressed in a recombinant form. We tested inhibitory activity in excretory-secretory products and homogenates of members family Diplozoidae towards coagulation factors IIa and Xa and their specific fluorogenic with 4 negative and 1 positive results. From the results of two transcriptome analysis we discovered three protein families of potential anticoagulants - annexins, serpins and Kunitz-domain proteins. For further analyses we focused on the Kunitz protein family. These proteins contain one or more structurally related active domains which are able to...
|
|
|
The impact of apixaban on overall hemostatic potential.
Cablíková, Ladislava ; Pávek, Petr (advisor) ; Carazo Fernández, Alejandro (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Bc. Ladislava Cablíková Supervisors: Ass. Prof. Mojca Božič-Mijovski, Ph.D., prof. PharmDr. Petr Pávek, Ph.D., RNDr. Jana Nekvindová, Ph.D. Thesis title: The Impact of Apixaban on Overall Hemostatic Potential Disorders at certain levels of the complicated haemostatic system can lead to either bleeding or excessive blood coagulation. These pathological conditions are treated with anticoagulants, which aim to correct excessive coagulation. However, traditional anticoagulant therapy has many limitations, which initiated efforts to develop oral anticoagulants with a better profile. These new-generation anticoagulants are called DOAC - Direct Oral AntiCoagulans. Apixaban, as one of xabans, has predictable pharmacokinetics and pharmacodynamics and therefore does not require a routine laboratory monitoring of the treatment effect. Nevertheless, it still requires evaluation in urgent clinical situations. Standard coagulation screening assays, e.g., PT (prothrombin test) and APTT (activated partial thromboplastin test), do not fully reflect the actual status of the drug. Therefore, researchers aim is to find a relatively simple and fast hemostatic assay that would correlate with the actual condition...
|
|
|
The impact of apixaban on overall hemostatic potential.
Cablíková, Ladislava ; Pávek, Petr (advisor) ; Carazo Fernández, Alejandro (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Bc. Ladislava Cablíková Supervisors: Ass. Prof. Mojca Božič-Mijovski, Ph.D., prof. PharmDr. Petr Pávek, Ph.D., RNDr. Jana Nekvindová, Ph.D. Thesis title: The Impact of Apixaban on Overall Hemostatic Potential Disorders at certain levels of the complicated haemostatic system can lead to either bleeding or excessive blood coagulation. These pathological conditions are treated with anticoagulants, which aim to correct excessive coagulation. However, traditional anticoagulant therapy has many limitations, which initiated efforts to develop oral anticoagulants with a better profile. These new-generation anticoagulants are called DOAC - Direct Oral AntiCoagulans. Apixaban, as one of xabans, has predictable pharmacokinetics and pharmacodynamics and therefore does not require a routine laboratory monitoring of the treatment effect. Nevertheless, it still requires evaluation in urgent clinical situations. Standard coagulation screening assays, e.g., PT (prothrombin test) and APTT (activated partial thromboplastin test), do not fully reflect the actual status of the drug. Therefore, researchers aim is to find a relatively simple and fast hemostatic assay that would correlate with the actual condition...
|
|
|
Anticoagulation factors and blood uptake by monogeneans of the family Diplozoidae
Skipalová, Karolína ; Mikeš, Libor (advisor) ; Sojka, Dan (referee)
For the successful food intake by organisms that feed on blood is essentials presence of antihaemostatic molecules such as vasodilators, anticoagulant molecules and apyrases., Although members of family Diplozoidae (Heteronchoinea) are blood-feeding parasites on the gills of the fish, these molecules, that could disrupt host hemostasis, have not yet been identified. Thus, the aim of this study was to find molecules with potential anticoagulant activity in homogenates of whole worm bodies and excretory/secretory products of the members of family Diplozoidae. Furthermore perform bioinformatics analysis of sequences obtained from transcriptom project of Eudiplozoon nipponicum (Heteronchoinea: Diplozoidae) and selected proteins (protein domain) then expressed in a recombinant form. We tested inhibitory activity in excretory-secretory products and homogenates of members family Diplozoidae towards coagulation factors IIa and Xa and their specific fluorogenic with 4 negative and 1 positive results. From the results of two transcriptome analysis we discovered three protein families of potential anticoagulants - annexins, serpins and Kunitz-domain proteins. For further analyses we focused on the Kunitz protein family. These proteins contain one or more structurally related active domains which are able to...
|
|
|
Different approaches to genetic testing of Leiden mutation
HÁJKOVÁ, Markéta
This thesis is about various approaches to genetical testing of Factor V Leiden. Factor V Leiden is currently one of the mostly occured hereditary causes of thrombosis. It's examination is one of the mostly required among the thrombophilic mutations. Thrombophilic conditions are serious health complications when most oftenly deep venous trombosis occurs, which means formation of blood clot in deep vein system of legs. This condition may cause health complications especialy during gravidity, but it may have serious consequences even for healthy person. If examination for Factor V Leiden is recommended by medical specialist then it is paid from the health insurance otherwise it is payed by person who requests the examination. Purpose of theoretical part of the thesis is to offer information about physiology of hemostasis and to understand mechanism of this system, further is about hereditary of Factor V Leiden and associated hazards and about rules of indication of genetic examination of this disorder. There is also description of risks related with use of hormonal anticonception and formation of thrombophilic conditions. And lastly this thesis concernes about various genetical methods which are useable for detection of factor V Leiden. Practical part consists of procedures of genetical methods and obtained results. Used methods were RFLP-PCR, real-time PCR and reverse hybridization on the stripes. Part of the results is statistical processing of different concentrations of DNA obtained by isolation from peripheral blood or from buccal swab and statistical overview of factor V Leiden identifications throughout the years in laboratories of company GENLABS s.r.o. Carriers of this mutation have increased risk of occurrence of thrombosis which can cause serious health conditions like pulmonary embolism or myocardial infarction. Indivinduals with such mutation can be heterozygotes aswell as homozygotes. Heterozygotes have five to ten times higher chance of mutation occurance then healthy person but homozygotes have this chance eighty to one hundred times higher. In human population factor V Leiden is spread unequally. In white population is estimated occurrence around 15% on the other hand occurrence on humans with African or Asian descent is very rare.
|
|
|
Anticoagulation factors of haematophagous parasites
Skipalová, Karolína ; Mikeš, Libor (advisor) ; Kolářová, Iva (referee)
2. Abstract Presence of antihaemostasis molekule is essential for the successful reception of food organisms that feed on blood. They are vasodilators, anticoagulant molecules and apyrases that break into the host hemostasis, thus blocking the whole process. This work summarizes all available information about the anticoagulant factors of haematophagous parasites. The blood sucking groups of helminths, arthropods and leeches is particularly inhibitors of serine proteases, which have their inhibitory aktivity directed primarily against thrombin and factor Xa. These are two key factors in the coagulation cascade. Factor X is the first member, where it sets the intrinsic and extrinsic coagulation pathways. At the same time it arises due to the aktive form of the protein thrombin, which is responsible for the conversion of soluble fibrinogen to insoluble fibrin, which stops bleeding. In addition to these two factors are inhibited of other members of the cascade. For example the inhibition of komplex factor VII and TF, which has been described in ticks and hookworms. Anticoagulation factors play a crucial role in the transmission of pathogens by blood from the vector into the host. The longer the blood is a liquid, the greater the chance of transmission of the pathogen.
|
|
|
Determination of coagulation factor VIII in haemophilia A patients;Evaluation of different methods of measurement depending on the type of mutations in the gene for factor VIII.
HOUSKOVÁ, Kateřina
Hemostasis is essential to life; it is the ability of organism to stop bleeding and to maintain the fluidity of the blood in an intact vascular bed at one time. Factor VIII is called antihemophilic globulin or antihemophilic factor. It is a plasma glycoprotein composed of two noncovalently associated chains. One chain is a heavy one, comprising domains A1- A2- B and the light chain composed of the domains A3- C1-C2. FVIII activity was measured in two ways from historical reasons a) a one-phase method, where a mixture of FVIII deficient plasma and patient plasma is analyzed using APTT assay; the absence of FVIII in the patient plasma leads to lengthening of the time, b) two-phase method, where the first step leads to formation of FVIIIa and FXa and in the second phase there are thrombin and fibrin created. Two-phase method is difficult to implement in routine laboratory, and therefore it was stopped using during time. This method was substituted by the chromogenic method after development of the chromogenic substrates, where there is in the first step created FVIIIa and FXa in the presence of FIXa, phospholipids and Ca2+, and in the second step there is formed a yellow coloration by the addition of the chromogenic substrate. FVIII deficiency causes a severe bleeding disorder, hemophilia A. I performed measurements from May to September 2013 at Coagulation laboratory at the Institute of Hematology and Blood Transfusion in Prague, where I was employed. I examined in total 76 patients, hemophiliacs A, who were at least 8 days without any treatment or substitution. I assigned numbers to patient's samples to ensure anonymity of patients. I worked with the automatic coagulation analyzer STA- R Evolution? from Diagnostica Stago, which works on the principle of photometry and chronometry. I determined the factor VIII by one-phase method and two-phase method and I compared the results. The genetic part of the work was analyzed in the genetic laboratory, which is part of our department. I worked up the results obtained from both methods in 76 patients to the table and the graph. The group included 14 (18 %) moderate and 56 (74 %) of mild hemophiliacs, then 6 (8%) hemophiliacs who did not meet the criterion of a mild hemophilia A, but clinically they belonged into mild hemophiliacs. Based on the stated criteria, we found out that 15 (20 %) patients had a ratio of FVIII: C1st/FVIII: Chr or FVIII: Chr / FVIII: C1st 0.6, they differed significantly in their values set by one-stage clotting FVIII and FVIII set by the chromogenic method. A total of 11 patients with FVIII activity were higher in the single-phase method. At three patients FVIII: C1st was even on the upper limit of the normal value, while FVIII chromogenic method gave on average 16%. We managed to find a causal mutation in the FVIII in 14 patients with "the different results", we could not investigate 1 patient genetically because of the missing genetic material. Mutations in patients with lower activity of FVIII set up by the chromogenic two-phase method were concentrated predominantly into the A3 domain; mutations in patients with FVIII a lower activity set up by one-phase clotting assay were concentrated in the A2 domain. The results presented show, that diagnostic of any patient with mild or moderate hemophilia A should include determination of FVIII by both methods; FVIII: C1st and FVIII: Chr.
|
|
|
Dependence of derived fibrinogen on values of DH from APTT and QUICK(PT) methods
Klus, Michal ; Zapletal,, Ondřej (referee) ; Škutková, Helena (advisor)
Hemocoagulation, blood coagulation, is an important indicator of hemostatic balance in the human body. There are many ways how to investigate blood clotting. In practice, next to the tests investigating time of coagulation cascade from view of internal way (APTT – activated partial tromboplastin time) and external way (PT – prothrombin time) is often used determining of fibrinogen concentration by Clauss method. Derived fibrinogen method determined fibrinogen concentration, too, by subtracting form the clotting curve in PT test. The reaction for Clauss method is not necessary here. Derived fibrinogen is not used much in practice. This is the reason, why the thesis related to this project will try to find relationship between concentration of fibrinogen and standard tests APTT and PT. Clinical data will be used for this.
|
|
|
Anticoagulant therapy focusing on patients with atrial fibrillation
VANÍČKOVÁ, Martina
Thrombosis has recently become a widely discussed issue due to an increasing number of cases and problematic treatment. Once developed, thrombosis cannot be cured completely. Currently used anticoagulant treatment is limited to avoid complications such a hemorrhage or stroke. INR value checks (Quick tests) and monitoring covering at least 70% of the treatment period within the therapeutic range are necessary (2-3), but could be rather difficult to maintain in some patients. So-called new anticoagulants could prove themselves as an appropriate solution.
|
guest ::
