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New diagnostic and therapeutic approaches for the prevention of CVD in patients with familial hypercholesterolaemia
Altschmiedová, Tereza ; Češka, Richard (advisor) ; Rosolová, Hana (referee) ; Rašlová, Katarína (referee)
Atherosclerotic cardiovascular diseases are still the most common cause of death in Europe, despite new diagnostic methods and treatment. Although many accelerators of the atherosclerotic process are known, only LDL-cholesterol is considered to be the causal risk factor for atherosclerosis. Familial hypercholesterolaemia (FH) is an autosomal dominantly inherited disease whose carriers have had a high level of LDL-cholesterol since childhood due to reduced amount or function of LDL receptors . Determination of the causative mutation is not always possible and the diagnosis is established by using some scoring systems which take into account personal and family history and some typical signs (e.g. tendon xanthomas) in addition to LDL-cholesterol value. The treatment of FH is lifelong and to achieve the LDL- cholesterol target, combination therapy (ezetimibe, modern biologic therapy) in addition to statins , is often necessary. However statins are always the mainstay of the treatment. By retrospective analysis of data from 1236 patients diagnosed with FH, we confirmed the cardiovasular risk of these patients is different depending on the presence of other risk factors. At the highest cardiovascular risk were individuals with combination of risk factors - high level of LDL cholesterol and total cholesterol, as...
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Freezing technology of bull sperm in relation to its survivability and fertilization ability
Doležalová, Martina ; Stádník, Luděk (advisor) ; Jiří, Jiří (referee)
The aim of optimalization the insemination doses production is to provide the highest fertilization ability of spermatozoa during the demanding proces of processing fresh semen and its subsequent cryopreservation. Temperature changes causes spermatozoa damage during the cooling and freezing. Spermatozoa is exposed to cold shock and many others limiting factors, which leads to cell death and therefore to decline of fertilization ability of thawed insemination doses. For increasing spermatozoa resistance, exactly the plasma membrane resistance against cold shock was fraction of egg yolk LDL cholesterol (low density lipoprotein) at various concentrations into the comercially produced diluents added. It is believed that LDL acts possitively to plasma membrane and helps to maintain the fertilization ability of spermatozoa after thawing. Following step in the proces of insemination doses production is slow cooling of diluted semen and equilibration, when the straws are store at cooling box for 30 minutes to 240 hours. This period is necessary to penetrate of certain diluent components into the spermatazoa also maintain the balance between their intracellular and extracellular concentration. Also important is subsequent freezing temperature gradient of insemination doses. The most suitable freezing method is based on computer controlled temperature decline in freezing chamber which allows the precise control of ice crystals formation that could tear and kill the cell.
During 2012 to 2016 was repeatedly collected semen from the group of breeding bulls (n = 27, Holstein and Czech Fleckvieh breed) at AI centre. Semen which fulfill the standard entrance conditions in first step was evenly into several parts divided. For dilution the three types of comercially diluents AndroMed, Bioxcell and Triladyl with and without LDL addition were used. Into the diluents AndroMed and Bioxcell the concentration of LDL 4 %, 6 % and 8% into the dilent Triladyl 6 %, 8 % and 10 % was added. Diluted semen was filled into the glass capillares with volume 0,1 ml and temperature +4 °C. Subsequently the sample was placed to cold bath (0°C) for 10 minutes. Then the volume of capillare with physiological solution (37 °C) was mixed and for next 120 minutes was incubate. The effect of cold shock to proportion of live spermatozoa was evaluated by using Eosin and Nigrosine staining technique during heat test of spermatozoa survivability after spermatozoa heating and after 120 minutes of incubation.
The more suitable semen diluents which provide the higher spermatozoa resistance against cold shock were AndroMed and Bioxcell. Together the possitive effect of LDL addition into the diluents to lower decrease of proportion of live spermatozoa during heat test was found (P<0.05). The most suitable LDL concentration which had a favorable influence at spermatozoa resistance against cold shock was 6 % in diluent Bioxcell. Values of the proportion of live sperm were higher at the beginning of the heat test (+1.31% to + 3.2%) and after 120 minute incubation (+5.82% to +8.41%) compared to other diluents with and without addition of LDL.
In the next step the process of equilibration was optimized, is an important part of insemination doses production. The effect of the length of equilibration for subsequent fertilization ability of spermatozoa was evaluated using spermatozoa motility based of CASA and proportion of live spermatozoa after thawing and during heat survival test lasting 120 minutes (37 ° C). Suitable semen was diluted by comercially used diluent AndroMed based on soya lecithin, filled into the straws (0.25 ml), cooled and equilibrated in cooling box for 30, 120 and 240 minutes and freezed in programmable freezing box applying four types of freezing curves differing in temperature rate decline. There was used standard and by producer recommended 3. phase freezing curve, then 2. phase freezing curve, and 3. phase freezing curve with slower as well as rapid decline of temperature rate in freezing chamber, compared with standard freezing curve. The highest spermatozoa motility was found using 240 minutes of equilibration by +2.72% and +4.58% compared to other lengths of equilibration (P <0.05 to 0.01). The highest proportion of live spermatozoa was found using 120 minutes of equilibration (+6.87 % and +8.68 %). The highest average spermatozoa motility during heat test after thawing was achieved by using 2. phase freezing curve (from +2.97% to +10.37%, P <0.05), also in the proportion of live spermatozoa (from + 4.37% to +8.82%, P <0.01). When evaluating interaction between the length of equilibration and freezing curve (standard 3. phase and 2 . phase freezing curve), the highest average spermatozoa motility and proportion of live spermatozoa using 240 minutes of equilibration by both freezing curves was reached, there was no statistically significant differences. As well as, in all evaluated parts of this study the individual differences between ejaculate of bulls and within semen from one bull (P <0.05) as secondary effect were found.
To maintain good fertilization ability of semen during cryopreservation is necessary to increase the spermatozoa resistance against cold shock using addition of correct concentration of LDL into the commercially used diluents AndroMed and Bioxcell. Subsequently the fertilization ability of insemination dose is influenced by cooling, the length of equilibration and freezing. The length of equilibration 120 minutes and more as well as gentle way of freezing according to freezing curve, which ensures a gradual decrease of temperature in freezing chamber provided the higher average spermatozoa motility and proportion of live spermatozoa.
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Ovlivnění plazmatické hladiny cholesterolu modelového organizmu polynenasycenými mastnými kyselinami n-3
Čeganová, Markéta
The aim of this thesis is to assess the influence of the polyunsaturated fatty acids, especially DHA and EPA in the diet on the animal organism in accordance to the levels of total HDL and LDL cholesterol and triglycerides in plasma. As experimental animals were used rats and they were divided into 3 groups of 10 pieces: control group with the addition of 6 % Carthamus tinctorius (n-6 flares), the group with the addition of 6 % fish oil (n-3: EPA, DHA fish) and the group with 6 % addition of oil from Schizochytrium (n-3: DHA, DHA). After 40 days the animals were putted to death according to relevant regulations. Blood lipids were obtained from blood samples by spectrophotometry. Value of polyunsaturated fatty acids of n-3 and n-6 was set by gas chromatography in the liver, muscle and adipose tissue. In accordance to the increase of content n-3 PUFA in group with fish oil and with oil from Schizochytrium, total and LDL cholesterol was reduced in comparison of samples in control group. In comparison to the control group were TAG reduced only in the group with addition of oil.
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Knowledge of plasma lipids of university students, smokers
VARAUSOVÁ, Anežka Gabriela
Introduction: Cardiovascular diseases are among the most common causes of death. The risk factors include, among others, hypercholesterolemia. It has been long believed that a high level of cholesterol intake via meals (dietary cholesterol) has negative effects on the cardiovascular system and therefore it has been deliberately removed from diet. Several studies, however, confirm the assumption that cholesterol in diet (dietary cholesterol) has little effect on cholesterol levels in blood. The human body receives cholesterol through nourishment in form of animal fats, but it can produce it in the liver, too. So if cholesterol is removed from nourishment due to a diet, the liver can cover the need of cholesterol through increased production. Cholesterol level in the human body is influenced by many factors most of which we can control. These factors include food containing too much saturated fat or cholesterol, overweight, physical inactivity, congenital health conditions, age and gender. There are no symptoms of hypercholesterolemia at the first stage and that is why the current population does not pay such attention, which it should deserve, to hypercholesterolemia. Because there are a number of educational materials on this subject, I decided to explore among the university students (especially smokers) if they are interested in this subject and what their knowledge is. Objective: The main goal of my thesis was to map out the university students´ (smokers) knowledge of plasma lipids. The bachelor thesis was prepared by means of a quantitative research using questionnaires. The research sample consisted of 300 full-time students. I collected 270 filled-in questionnaires out of the 300 distributed. The questionnaire return rate was therefore 90%. 99 out of those 270 were unusable due to the fact that the respondents did not belong to the target group. The total number of questionnaires to be evaluated was 171 questionnaires. Results: I defined a research hypothesis. Hypothesis # 1 Knowledge of plasma lipids of university students, smokers is inadequate, has been confirmed. 36 (21%) respondents out of the total 171 respondents knew their level of cholesterol and 135 (79%) of the respondents did not know their level. Conclusion: This thesis can be used in practice as a publication, to increase the general awareness of students and professors of the issue of cholesterol. An educational material is attached to this thesis that can be used in general practitioners? surgeries.
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Influence of increased aplication of the file polyunsaturated fatty acids on alteration of level cholesterol of individuum.
FLANDERKOVÁ, Michaela
I have engaged in the influence of increased aplication of the file polyunsaturated fatty acids on alteration of level cholesterol of individuum in my bachelory work. At first I had studied theoretical knowledge focused the theme of polyunsatureted fatty acid and cholesterol. I had devoted to this knowledge in theoretical part of my bachelory work. After I had contacted some people. I had known that they have increased cholesterol level. This people I had acquainted with themes of my bachelory work and development of research. The acquainted people have accepted part of research. Initial measuring taked place after. The measuring determined the level of total cholesterol, LDL cholesterol, HDL cholesterol, triacylglycerol. The three month interventionist program taked place after, from march 2011 to december 2012. The Participant of research had consumed of food supplement, which contented fish oil. Fish oil is rich in content of polyunsaturated fatty acid omega 3 (EPA and DHA). Output measuring taked place after three month. On the basis of measuring got results. I worked with results in the practical part of my bachelory work. I have compared results of initial and output measuring.
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Laboratory markers in the process of extracorporal elimination of lipoproteins in long-term treatment of severe familial hypercholoesterolemia
CHOCHOLOUŠOVÁ, Jana
LDL-apheresis is a method of extracorporeal LDL-cholesterol elimination, which is known to decrease inflammatory processes of atherogenesis. The study is divided into two parts. THE AIM of the first part was to evaluate LDL-apheresis procedures based on immunoadsorption in 9 patients, 5 men and 4 women, (median age was 57, range 19-61 years), with severe familial hyperlipidemia (period 2004-2007). Immunoadsorption was performed by means of Cobe Spectra (COBE BCT, USA), ADA or Adasorb (Medicap, Germany) and columns of LDL-Lipopak (Pocard Ltd., Russia). Before and immediately after the procedure, blood samples were taken and serum levels of lipoproteins and some other parameters were determined. The aim of the second part was to evaluate the effectiveness of rheopheresis method and compare it with LDL-apheresis. Procedures of rheopheresis were implemented in 4 patients, 3 men and 1 woman, median age was 51 (35 {--} 67) years. During this procedure Cobe-Spectra was used; plasma was than pumped into the secondary grade filtrs Evaflux. Before and immediately after the procedure, blood samples were taken and serum levels of lipoproteins and some other parameters were determined.
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