The aim of the diploma thesis was measurement and characterization of EEM (excitation emisssion spectra) of IHSS humic acids standards and soil, lignite humic and fulvic acids and humates prepared from different sources. The aim was to follow the differences in their fluorescence spectra and to determine precise location of fluorescence peaks. Target of this work was to find the optimal way for exporting of scan data to the 3D EEM spectrofluorographs. The dependence of relative intensity of fluorescence on pH value was studied further.

This work deals with the study of the possibility of creating an electrochemical biosensor for the detection of specific nucleic acid sequences. The theoretical part describes various types of RNA, with a special focus on miRNAs and possibilities of its detection. It also describes various electrochemical detection approaches, both nucleic acids, and more specifically miRNAs. miRNA is a potential biomarker that can be used for early and non-invasive diagnosis of cancer. The experimental part is aimed at designing, optimizing and preparing a sensitive biosensor for the detection of miRNA-21. A biosensor was prepared to detect the lowest concentration of miRNA-21 in the sample. A linear concentration range for the calibration curve from 1 nM to 1 µM concentration was measured. We measured LOD of 1 fM and by calculation from the regression equation to 3.2 zM and LOQ was 10.8 zM. Subsequently, samples were measured in artificial urine to verify the functioning of the sensor in real conditions. The results showed a minimal effect of the matrix on the determination of the target miRNA.

When considering fish as food, first of all the n-3 LC-PUFA, particularly EPA and DHA are in focus. Furthermore, it gets obvious that the proteins and peptides in fish have not only a high nutritional value but also impact on human health issues. In addition, fish can be considered as a good source of several minerals, vitamins and micronutrients. The overall aim of the thesis was to highlight different factors which influence nutritional quality of fish and to focus on the nutritional value of some commercially important river fish species from the Czech Republic. Moreover, to examine different sustainable alternatives to replace FO and FM in the feed of carnivorous. According to our results, there were some variation of FA composition in the selected seven freshwater fish species from the Czech Republic, depending on the natural habitat and differences in feed and its availability. Simultaneously, we observed a very favourable FA composition with good proportions of n-3 PUFA, including EPA and DHA in all analyzed species which reflects the composition of the natural diet. Consequently, the values of both index of atherogenicity (IA) and index of thrombogenicity (IT) were low and close to the values of the so-called Eskimo diet, which is related to very low incidences of the coronary heart disease. According to our findings we concluded that the chosen species have a standard protein content, minimum carbohydrates and relatively low contents of fat, which can, however, vary to some degree in various localities, most probably related to the availability and composition of the feed. Due to the combination of the drastic increase in the need for aquaculture feed as well as decline in the sources of FM and FO, utilization of alternative sources received more attention. Based on our result, it is possible to replace VO by yeast oil produced from lingocellulose in the feed of Arctic char (Salvelinus alpinus). There were no significant differences in the study, regarding weight gain, feed conversation ratio, condition factor and hepatosomatic index between the control and the yeast oil fed group. According to the results of another study, partial (25% of pellet replaced live house cricket, 25% of pellet replaced by superworm, combination of 12.5% crude energy of each group) or total replacement (50% of each) of house cricket and superworm for FM in the commercial diet of rainbow trout indicated no negative effects on growth, survival, FCR and gross energy utilization. Inclusion of insect was connected with lower content of nutritionally valuable n-3 FAs (EPA and DHA). In our study total replacement showed the mixture of insects, caused the better growth performance compared to the commercial feed of similar energy value as insects are good live food for salmonids. With increase in the proportion of insect's inclusion in the feed of fish, changes in the sensory properties, texture and colour of fish flesh was in a way that showed less acceptability and preference by consumers. Replacement of 25% FM by a mixture of insect meal including house cricket- (Acheta domesticus) and superworm- (Zophobas morio) meal (with an amino acid adjustment) in the diet of perch, on survival, growth, feed conversion with special emphasis on lipid changes and composition showed FA composition of the fish fillet was only affected to a minor extent. However, the lower growth performance in the fish group fed by insect indicated lower nutritional value and digestibility of the feed along with the taste of the feed. Interestingly, the significant increase of 18:2 n-6 in the group fed by insect was due to the higher content of this FA in both insects and subsequently in the experimental diet which from the nutritional point of view this change was neglectable.

Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Student: Marie Šimková Supervisor: Mgr. Alena Prašnická Title of diploma thesis: Alternation of bile production due to iron depletion Introduction: Liver has an irreplaceable role in the production and secretion of bile. This body fluid serves as the main excretion way of some endogenous and exogenous substances. Another liver property is the ability to store substances essential for correct functions of the body, e.g. iron. It has been shown that iron could have an impact on the bile production and secretion. Aim: The aim of this diploma thesis was to discover an impact of iron depletion on the bile synthesis and metabolism, especially on bile acids, and the way it affects transporters expression. Methods: Male Wistar rats (n=6 in each group, 250 ± 20 g) were divided into two groups: control group (Chow diet) fed with standard diet and iron depletion group (ID), fed with iron depletion diet for 21 days. To investigate the changes in bile flow, the bile had been collected for 120 min during in vivo clearance study. The analysis of the changes in expression of bile transporters and enzymes responsible for de novo bile acid synthesis was performed at the mRNA (qRT-PCR) and protein (Western blot)...

The aim of the study was to study the variability of the gene in the population FABP3 pig breed Czech large white. Analyze their association with indicators of pork production. Gene FABP3 belonging to multigene families FABP (fatty acid binding proteins affecting the transport of long chain fatty acids). Analyzed polymorphism was at position X98558:g.1321G>C in the 5'UTR region. The relative frequencies of genotypes were: HH = 0,87, Hh = 0,13. hh genotype was not detected in the monitored individuals. The values of the relative frequencies of alleles were as follows: H = 0,93 h = 0,07. Based on the association analysis showed a statistically significant difference between genotypes for stearic acid and linoleic acid. Highly statistically significant difference was observed with palmitic acid.

The aim of this work was demonstrate positive effect of n-3 polyunsaturated fatty acids EPA and DHA on the concentration of plasma triglycerides and cholesterol. According to the hypothesis, these polyunsaturated fatty acids are capable of reducing plasma cho-lesterol in mechanism, where is increased gene expression Insig-1 and simultaneously reduces the expression of genes encoding an enzyme for cholesterol synthesis (3-hydroxy-3-methylglutaryl-CoA reductase, a gene for HMG-CoA-R) and the gene for the LDL receptor, which is responsible for the entry of cholesterol from the plasma into the cells (low density lipoprotein receptor, the density, the LDL-R gene). LDL-R.The experiment was conducted on a model animal (Rattus norvegicus) fed with a standard feed mixture with the addition of 3% of salmon oil, fish oil 6% and 6% of oil from algae of the genus Schizochytrium. The control group was fed a feed mixture with the addition of 6% safflower oil, food with the addition of 3% palm oil was used as a negative control. Each group consisted of 10 animals. From the blood samples was determined levels of total cholesterol, HDL-cholesterol, LDL-cholesterol and triglycerides. The fatty acid content was determined in muscle, liver and adipose tissue. Food enriched with eicosapentaenoic acid and docosahexaenoic acid led to a reduction in total cholesterol and LDL-cholesterol levels. It was shown (P <0.05) after the addition of fish oil, salmon oil and algae of the genus Schizochytrium oil to feed mixtures of experimental rats, a decrease in total cholesterol (27%, 21%, 27% respectively) relative to controls. Our attempt partially confirmed the hypothesis. At the same time, we found that plasma lipid metabolism affects different mechanism than we thought.

Pork represents the largest volume of meat production in terms of the global market. In the Czech Republic accounted for over 50 % of total meat consumption. Pork is often cited as a food with a high level of fat and unfavorable balance of fatty acids. Intake of dietary fats should contain saturated (SFA), monounsaturated (MUFA) and polyunsaturated (PUFA) fatty acids in a ratio of <1 : 1,4 : >0,6. The PUFA/SFA ratio should be higher than 0,4 and the n-6/n-3 PUFA ratio should be 5 : 1. To increase awareness of the importance of food to human health has recently been an effort to find ways to produce healthier meat, i.e. with a higher proportion of PUFA to SFA and better ratio of n-6 and n-3 PUFA. Pork quality in terms of its composition of fatty acids is currently not very favorable. Fatty acid profile in animal fats can affect by the fat contained in food and change the composition of fatty acids in the fat of pigs through the nutrition. The diploma thesis describes the fats, their importance in human nutrition, a group of fatty acids found in fats, their sources and factors influencing the content and fatty acid profile in muscle and adipose tissue of animals. The aim of the study was to investigate the influence of additions of rapeseed and soybean oil, as a major source of unsaturated fatty acids, in pig fattening on the quality and composition of intramuscular fat (IMF) in the pork loin (MLLT). It was also evaluated the effect of the addition of these oils on selected parameters of fattening capacity and carcass value. For a more objective evaluation of the suitability of fat was calculated atherogenic and thrombogenic index. The experiment was included 32 pieces of slaughter pigs for in the final hybrid combinations DanBred. Pigs were throughout the fattening fed ad libitum complete feed mixtures and depending on the feeding divided into three groups, i.e. control group without the addition of oil and an experimental group containing 4% of rapeseed oil and an experimental group with the addition of 4% soybean oil. The results show that the group with the addition of soybean oil exhibited simultaneously with the lowest daily intake of feed the best feed conversion. The addition of rapeseed and soybean oil had no significant effect on the physical parameters of the pork loin and backfat. So, technological quality of backfat has not been influenced. Amount of IMF not influenced by any of the oils. However, fatty acid composition in IMF has been influenced, both of oils. Although, SFA has not been influenced neither one of the oil, content of the most represented SFA, palmitic acid, was significantly reduced in the soy diet. Significantly lowest content of MUFA showed group with soybean oil, which was due to a significant reduction of the most represented acid, oleic acid. Treatment with the addition of both oils were significantly increased PUFA and n-3 PUFA. The PUFA/SFA ratio was also increased, the most by soybean oil. The ratio of n-6 / n-3 PUFA was significantly reduced by the rapeseed oil. Content of linoleic acid, alpha-linolenic acid and eicosapentaenoic acid was significantly increased as rapeseed and soybean oil. Soybean oil increased content of alpha-linolenic acid significantly more than rapeseed, which against significantly increased linoleic acid. Thrombogenic index was significantly reduced by rapeseed and soybean oil, atherogenic index only by soybean.

This paper deals with a comparison of the profile of fatty acids in adipose tissue pigs with regard to the differently enriched feed of unsaturated fatty acids. In the theoretical part, the issue is handled using professional literature, supplemented by chapters dealing with the digestive system and digestion. In the practical part, the experiment was conducted. The experiment was performed at the pig breeding test station at Ploskov near Lány. The experiment included 72 pieces of slaughter pigs of balanced sex (36 barrows and 36 gilts) final hybrid combinations DanBred. This paper presents a methodology, under which the test was performed. The pigs were included in the test in the average weight of 29.2 kg and the average age of 70 days. The pigs were fed with a complete feed mixture. The pigs were according on diet divided into six experimental groups with supplemented 4% oil (rapeseed and soybean) and one control group without added oil. In experimental groups were incorporated oil into the feed mixture, for a period of 6, 4 and 2 weeks prior to slaughter. Age and average slaughter weight of pigs at the end of the fattening period was 152 days and 115.8 kg. From the measurement results, it can be stated that the resulting interaction between the time of application and the type oil used by the addition of has been found in the content of palmitic acid, linoleic acid and alfa linolenic acid, n3 PUFA and n6 PUFA and the ratio of n3 and n6 PUFA and also in oxidation stability, carried out immediately after thawing. Finally, it is possible to say that the profile of fatty acids contained in the feed is reflected in the profile of fatty acids in adipose tissue. When the rapeseed oil was fed the content of MUFA increaset and when the soybean oil was fed the content of PUFA increaset in the adipose tissue. Time of feeding fatty acids had no content great influence. Thus, from economic point of view the best option seems to be the addition of soybean oil in the diet two weeks before slaughter.

The two ecotypes of Noccaea caerulescens originating from two differently contaminated sites (Redlschlag, Austria; Ganges, France), N. praecox (Mežica, Slovenia), Arabidopsis halleri and Spinacia oleracea were studied to compare Cd accumulation and tolerance. After 30, 60, 90 and 120 days of cultivation in Cd contaminated soil (30, 60 and 90 mg Cd/kg soil) for hyperaccumulators and 25, 40, 55 and 75 days of cultivation in Cd contaminated soil (30, 60 and 90 mg Cd/kg soil) for spinach gas-exchange parameters (net photosynthetic rate, transpiration rate, stomatal conductance, intercellular CO2 concentration), fatty acids, macro- and microelements were determined as well as N utilization by plants. Noccaea species and A. halleri showed similar changes in yield of biomass, Cd content and total content of amino acids. These species confirmed differences in the profile and contents of individual free amino acids. The comparison between Noccaea species showed that Cd stress resulted in similar changes of gas-exchange parameters. Contrasting responses of plants to Cd contamination were confirmed in macro- and microelements contents, fatty acids and amino acids metabolism. Spinach showed contrasting response to cadmium compared to hyperaccumulators, especially in profile of free amino acids and fatty acids.

Human nutrition is a function of nutrients' intake from food. The essential nutrients needed by the human body to produce energy, growth and repair of cells, tissues and organs and which accepts in the human diet, are fats. They are obtained from plant or animal sources. Especially animal fats are often ranged among the unpopular food components. Czech Republic is a country with traditionally high consumption of pork. Recently, about healthy foods greatly increase the interest. These include some animal products having an altered composition of fatty acids in fats. The content and composition of saturated and unsaturated fatty acids in human nutrition are important for protecting of human health. The high content of saturated fatty acids in foods and unfavorable ratio between the n-6 and n-3 fatty acids may be the cause of many diseases, especially cardiovascular diseases. Fats in the diet should contain saturated, monounsaturated and polyunsaturated fatty acids, in a ratio of about 1: 1.4: 0,6. From a health perspective, the most important are the n-3 polyunsaturated fatty acids. Animal fats, however, contain sufficient n-6 polyunsaturated fatty acids, n-3, however, a little. Pork meat quality from the viewpoint of fatty acids composition is currently not very favorable. Significant effects on the fatty acids profile of animal fats have the fats in the feed. It is thus the possibility of by using the diet to influence the composition of fatty acids in the fat in pig. The work is focused on the possibility of influencing the fatty acids composition of pork intramuscular fat using supplementation of different sources and different feeding techniques of unsaturated fatty acids in the feed in pigs. In the experiment, as a source of unsaturated fatty acids the supplement of the rapeseed or soybean oil was used. The results demonstrate that the above unsaturated fatty acid sources significantly changes the fatty acid composition of intramuscular fat.