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Amylase gene variability in human populations
Vosmíková, Veronika ; Černý, Viktor (advisor) ; Daňková, Pavlína (referee)
Amylase is a starch-digesting enzyme produced by the pancreas and salivary glands in humans. Genes for the salivary isoenzyme (AMY1), located on chromosome 1p21.1, show remarkable variability in their copy numbers (CNV). Their count is positively correlated with the final amount of the produced enzyme. This variability could be the result of positive selection during human evolution, depending on the amount of starch consumed by various populations. Starch pre-digested by salivary α-amylase is absorbed more effectively, resulting in a greater gain of energy in later stages of the metabolizing process. Thus, it could have been one of the factors for rapid brain growth in humans and, significantly later, even for the agricultural transition. It could belong to a group of other known subsistence-based evolutionary changes in the human genome, such as those resulting in lactase persistence or slow acetylation of xenobiotics. Nowadays, the number of copies and the amount of produced α-amylase can influence the glucose level in the blood and insulin production after consumption, as well as the incidence of certain civilization diseases, such as obesity or type II diabetes. The influence of salivary α-amylase on the composition of the microbiome, cardiovascular diseases, and the pro-inflammatory profile...
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Global phylogeography of the deep-sea fishes
Knězů, Tereza ; Musilová, Zuzana (advisor) ; Vukićová, Jasna (referee)
Phylogeography of deep-sea fish remains poorly explored, largely due to the extreme conditions in which these organisms live and limited technological capabilities for researching these species. The aim of this thesis is to summarize the information known about the phylogeography of deep-sea fish, identify factors influencing population distributions, and connect them on a global scale. The distribution of deep-sea fish populations is influenced by physical, topographic, and hydrographic factors. Their connectivity is driven by the migratory abilities of fish and the utilization of marine currents for their dispersion. Migration is divided into vertical and horizontal. The vast majority of deep sea fish undergo some form of migration, at least during ontogenetic development. The most common is diurnal vertical migration, where fish regularly move to shallower depths at night for feeding. Molecular methods, mainly mitochondrial DNA markers, were used for research to determine the phylogenetic tree of species. The results suggest that the topography of the seafloor is rarely a barrier to the flow of genetic information. Furthermore, the results often refute the hypothesis of isolation caused by the distance between populations. They often exhibit a relatively high rate of panmixia. When speciation...
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Western blot analysis of markers of endothelial dysfunction in mice aorta following high-fat diet administration
Střelcová, Ivana ; Němečková, Ivana (advisor) ; Vitverová, Barbora (referee)
7 2. ABSTRACT Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Thesis: Western blot analysis of markers of endothelial dysfunction in mice aorta following high-fat diet administration Supervisor: RNDr. Ivana Němečková, Ph.D. Author: Ivana Střelcová Background: High dietary fat intake is associated with increased risk of developing various metabolic diseases, including increased cardiovascular risk. The aim of this thesis was to investigate whether the administration of the high-fat diet containing 42 % kcal of fat affects expression of selected markers of endothelial dysfunction, the key mechanism in development of cardiovascular complications. Methods: In this thesis were evaluated changes in the expression of endothelial nitric oxide synthase (eNOS), endoglin (ENG), endothelin-1 (ET-1), intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1). An experimental group of male C57BL/6J mice aged 7 months was fed the high-fat diet containing 42% kcal of fat for 12 weeks. The control group was fed a standard laboratory diet. Western blot analysis followed by detection using a chemiluminescent reagent was used to evaluate the observed markers of endothelial dysfunction Results: The results show that there were no...
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Proteomic resolution of cereals
Nyškovská, Kristýna ; Kučková, Štěpánka (advisor) ; Kolář, Karel (referee)
This bachelor thesis deals with the proteomic differentiation of cereals (amaranth, sown barley, sown maize, qinoa, grain sorghum, sown oats, sown wheat, sown buckwheat, sown millet, sown rice, spelt, sown rye). Cereals are abundant in the human diet and are one of the main sources of protein and carbohydrates. The aim was to determine whether it is possible to distinguish between different types of cereals and to compare the difference between cereals and pseudocereals based on the protein composition of their grains using the mass spectrometric method MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization - Time of Flight. The theoretical part deals with the introduction of cereals, pseudocereals and their nutritional values, application and analytical method MALDI-TOF MS. The experimental part deals with the collection of different cereal species and their subsequent analysis. The samples were mechanically processed and digested using the enzyme trypsin. The cleaved peptides were measured by MALDI-TOF MS and the obtained spectra were subsequently processed using mMass and PostgreSQL (version 14.2). In this way, specific m/z values were determined that are characteristic of each cereal analyzed. From these values, a table was compiled that summarizes these characteristic values. Which allows the...
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Vliv hlaváče černoústého (Neogobius melanostomus) na potravní řetězec řeky Ploučnice
STANĚK, Pavel
Biological invasions are prominent factor affecting changes in invaded ecosystems. Invasive species can alter in food web of the indigenous community, which leads to changes in the community through ecological and economic effects. This work was focused on the effect of the Round goby (Neogobius melanostomus, Pallas 1814) food web of river Ploučnice. Round goby was first recorded recently in the Czech Republic (2015). Thus, it can still be considered a young invader into Czech inland waters. The main goals of this thesis were: I) to reveal Round goby food source preferences, II) to Competition between Round goby and European bullhead (Cottus gobio, L 1758), as this species is a native endangered species present at this locality. III) Possible preferences of Round goby as a putative food source of predatory fishes in sampled localities. For this purpose, River Ploučnice has been chosen. River Ploučnice flows into Elbe River, which is fully invaded by Round goby. Thus, the bottom part of the Ploučnice River is invaded by Round goby as well. However, thanks to the presence of migratory barriers between localities, Round goby has not yet been detected in localities upstream. To reveal mention goals all biota units in all localities were sampled. Later, stable isotope analysis (carbon and nitrogen) followed by Bayesian models were employed. In general, results suggest that this species had the highest abundance among other species in localities with an established Round goby population. In addition, results reveal Round goby as a true generalist regarding food source preferences. Comparing trophic niche width and overlap showed a significant overlap between some of the ontogenetic groups of round goby and European bullhead, reaching up to 67,8 %. Also, increased niche overlap between ontogenetic stages of European bullhead was observed when Round goby was present, in comparison to localities without the presence of Round goby. Additionally, predatory fish food preferences showed no specialization on round goby as a food source, although round goby was part of their diets. In conclusion, this was found round gobyas a consumer, competitor, and even as a food source for predatory fishes.
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Od trusu k prostřenému ubrusu. Molekulární analýza potravy raka signálního
ŠKRABÁNEK, Jan
This thesis aimed to determine the diet of the signal crayfish in Křesánovský brook in 2020. In addition to that, we aimed to find and test suitable markers for the diet analysis of signal crayfish, which are described in the thesis. We analyzed the crayfish diet by examining DNA from faecal pellet samples collected monthly from April to September. By using group-specific DNA markers, we targeted the main dietary groups such as Heptagenidae (markers Hpt28Sf + Hpt28Sr), Ephemeroptera (Epa28Sf + Epa28Sr), Integripalpia and Spicipalpia (Inspi18Sf + Inspi18Sr), Plecoptera (Ple-gen A269 + Ple-gen S268), Chironomidae (Chiro18Sf + Chiro18Sr), and Gammaridae (Gamae28Sf + Gamae28Sr). We also focused on representatives of macrophytes Characeae (CHA677 + CHA1681R) and Eudicotyledons (MYR152F + MYR1696R). The results showed that the crayfish's diet varied during the sampling season. Gammaridae was the most common food item, found in 54.1% of the samples of faecal pellets. Heptagenidae, Integripalpia, and Spicipalpia were also significant, present in 31.3% of the samples. Plecoptera appeared in 20.8% of the samples, while Chironomidae and Characeae were found in 10.4%. Eudicotyledons were detected in 8.3% of the samples. Ephemeroptera were the least common food group, detected only in 2.1 %. Gammaridae were the most significant component of the crayfish diet in April (found in 7 samples), while Heptagenidae were the second most abundant component. The diet was more diverse in May, but Gammaridae remained the primary food source, followed by Integripalpia and Spicipalpia (in 5 samples). In June, Gammaridae was present in all samples, while Heptagenidae and Integripalpia/Spicipalpia were significant. In July, Heptagenidae was the most common food group (found in 5 samples), with Gammaridae found in 4 samples. No food items were detected in the samples from August and October, and only one sample contained Gammaridae in September. Overall, this study provides insights into both the seasonal variations in the diet of signal crayfish and markers suitable for its analysis.
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Host-microbe interactions and its consequences for intestinal inflammation and carcinogenesis
Kejzlarová, Leona ; Kverka, Miloslav (advisor) ; Smrž, Daniel (referee)
A number of physiological and pathological processes, including the transition from chronic inflammation to cancer, are affected by commensal microbes. However, abundance of microbes and ability to produce active metabolites in the intestine depend on environmental factors, particularly diet. Microbes can influence this process in two ways, by producing genotoxic substances that directly damage the epithelium or by stimulating the inflammatory response. The aim of my thesis was to study the interaction among gut microbiota, diet and the immune system with the subsequent influence on the development of colorectal cancer (CRC) in an experimental mouse model. Animals were fed synthetic diets containing either normal amounts of animal protein (17%; KD) or elevated amounts of animal protein (51%; HPD) throughout the experiments. Two weeks after the diets were introduced, intestinal tumors were induced by administering azoxymethane (AOM) and inducing acute inflammation with 2% sodium dextran sulfate one week after AOM injection. At the end of the experiment I evaluated the number of tumors in the colon and the status of the immune response in the intestine, mesenteric lymph nodes and spleen. To study the effect of macrophages, a similar experiment was performed in animals with depleted macrophages using...
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Influence of makroelements from food on DNA and epigenetic profile
Veselý, Zdeněk ; Kouřilová, Xenie (referee) ; Brázda, Václav (advisor)
The macroelements contained in food have an important function for the human body. They are involved in several of biochemical reactions in the body and their abundance can prevent serious diseases. The theoretical part of the bachelor thesis describes the function of minerals in the human body, the function of DNA and epigenetic mechanisms such as DNA methylation or histone modification. The influence of nutrition and function of selected macroelements – sodium, magnesium, calcium, potassium on epigenetic modifications and on the stability of G-quadruplexes was described. The aim of the experimental part of this work was to study the effect of these substances on DNA structures in vitro and to prepare them experimentally for in vivo studies.
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Monitoring of Phytoestrogens Expression to the Milk and Milk Products
Watzková, Jarmila ; Rittich, Bohuslav (referee) ; Doležal, Petr (referee) ; Omelková, Jiřina (advisor)
Phytoestrogens are very intensively studied in recent years because they show different pharmacological effects on human health. For example, it was demonstrated that phytoestrogens reduce the risk of certain cancers (breast, prostate or colon), act as antioxidants in the body and affect the activity of certain enzymes. In addition, also it was demonstrated number of positive effects of soybean phytoestrogens on the health status of women in the menopause, such as reducing the risk of atherosclerotic cardiovascular disease or osteoporosis. Various technologically processed soy beans are important part of the feed rations mainly for high-lactating dairy cows, because they are a valuable source of the energy and high-quality protein, despite their negative effect on reproductive performance. Phytoestrogens consumed from the feed rations are hydrolyzed in the rumen of dairy cow and together with their metabolites are absorbed in the small intestine into the blood. They can be excreted from the body in urine or milk. From this perspective, cow's milk may become a source of phytoestrogens substances, especially equol in the human nutrition. On the other hand, some research results suggest possible potential risks associated with phytoestrogens intake in the diet on human health. At the some age (infants and young children) and group (vegetarians) categories may be dietary intake of phytoestrogens undiserable. According to most experts, a beneficial effect on human health prevail the potential risks. However, the risks associated with phytoestrogens intake in food can not be ignored and therefore phytoestrogens are still actual problem in the field of scientific research.
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Using different methods of DNA isolation of lactic acid bacteria in molecular biological methods
Chvalkovská, Eva ; Skoumalová, Petra (referee) ; Brázda, Václav (advisor)
This thesis focused on the probiotic bacteria, DNA isolated from these bacteria by three different methods and the effect of isolation on DNA identification using molecular biological methods. Probiotic bacteria are an important part of human intestinal tract. They have an important role in the function of the immune system due to adhesion to the mucosa of the intestinal flora. They create a inhostile environment for pathogens. Probiotic bacteria are commonly taken in the food like dairy products or food supplements. However, overuse of antibiotics is at risk of passing on the intrinsic resistance that probiotic bacteria have to the pathogenic bacteria. The intrinsic resistence they have to maintain the natural homeostasis of the intestinal tract. It is important to effectively identify risky probiotic bacteria that have the ability to transmit resistance to eliminate their presence in food and dietary supplements. Three methods of DNA isolation like phenol extraction method, magnetic particle isolation and commercial kit isolation were used in the experimental part. DNA was isolated from three dietary supplements, namely Biopron 9 premium, Linex forte and GS Lactobacily forte 21. The purity and concentration of the isolated DNA was detected spectrophotometrically. The presence of individual DNA strains in dietary supplements was confirmed by real-time polymerase chain reaction. The best method of isolation in terms of purity and concentration of isolated DNA was evaluated by RT-PCR and spectrophotometry using a commercial kit isolation method.
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