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Comparison of the effect of bisphenols on parental and differentiated H9c2 cells
Macáková, Barbora ; Jirkovský, Eduard (advisor) ; Macháček, Miloslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Barbora Macáková Supervisor: PharmDr. Eduard Jirkovský, Ph.D. Title of diploma thesis: Comparison of the effect of bisphenols on parental and differentiated H9c2 cells Bisphenol is compound largely found in products and environment around us. Its toxicity and danger for human health is still being studied, meanwhile direct influence of environmentally relevant doses was not discovered. These endocrine disruptors are affecting variousreceptorsin bodiesand effecton cardiovascularsystem isnotexcludedeither.The most extensively studied substance is BPA, which is gradually being replaced by its analogs, although these are not extensively researched to the same extent. In this work we focused on influence of bisphenol BP, M and S on differentiation of H9c2 cells, serving as a model with potential negative effects on heart development and its regeneration after damage. H9c2 cell line has an undifferentiated phenotype but can be easily directed towards differentiation under certain conditions Research was focused on impact of bisphenol on differentiated cells and its changes in the expression of selected differentiation markers (GATA4, Hand2, Tnnt2, Myog) (transcription factors). Set protocols were...
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Diversity of Epilobium and methods to explore their hybridization
Kuchyňková, Lenka ; Kaplan, Zdeněk (advisor) ; Šemberová, Kristýna (referee)
Willowherbs (Epilobium) belong to the genera whose evolution is significantly affected by interspecific hybridization. Although this process often generates evolutionary novelties and leads to speciation, it can also result in genetic erosion and extinction. This thesis is mainly focused on the Czech Epilobium species that are potentially endangered by interspecific hybridization. The emphasis falls on the group of rare mountain willowherbs including E. nutans, E. alsinifolium and E. anagallidifolium. In addition to frequent interspecific hybridization, the research of this group is also complicated by the subtlety of distinguishing morphological features. The review also summarizes the available methods that can be used to study interspecific hybridization in the follow-up thesis.
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Metabarcoding and environmental sequencing projects
Kandaurova, Ekaterina ; Hampl, Vladimír (advisor) ; Poláková, Kateřina (referee)
The thesis explores the use of metabarcoding in projects uncovering biological diversity and its applications in various fields of biology. The thesis provides an overview of the principles and workflow of metabarcoding, including the choice of specific molecular markers for various groups of organisms and the bioinformatics aspect of data analysis. Limitations of metabarcoding, such as lack of standardization, primer bias and targeting metabolically inactive organisms are discussed along with potential strategies for overcoming these obstacles. The practical application of metabarcoding is demonstrated on examples of its use in sequencing projects, such as Human Microbiome Project or TARA Oceans expedition. The potential of using metabarcoding for research purposes is discussed from the perspective of paleontology, diet analysis, and conservation biology. Overall, this work aims to assess the potential of metabarcoding as a powerful tool and could serve as an initial guidance for researchers interested in utilizing metabarcoding for their scientific investigations. Keywords: metabarcoding, eDNA, molecular markers, NGS, bioinformatics, biodiversity.
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Characterization of different differentiation protocols of H9c2 cardiomyoblasts
Gardianová, Anna ; Jirkovský, Eduard (advisor) ; Macháček, Miloslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Anna Gardianová Supervisor: PharmDr. Eduard Jirkovský, Ph.D. Title of diploma thesis: Characterization of different differentiation protocols of H9c2 cardiomyoblasts Cell differentiation is a common process in the development of multicellular organisms leading to an alteration of cell's appearance and function. The process of differentiation plays a significant role in the reparation of damaged tissues too. Some chemicals may have negative effects on these processes and therefore affect human health. This thesis focuses on characterization of differentiation processes of H9c2 cell line using various published differentiation protocols, and to describe changes over time in the morphology (occurrence of multinucleated and elongated cells) and mRNA expression of selected markers typical for the cardiac (cTnT, Hand2, GATA4) or skeletal (myogenin) muscle phenotype. We monitored the process of the differentiation for 15 days and the individual protocols differed by the addition of fetal bovine serum (FBS, 1 % or 10 %) and all-trans- retinoic acid (ATRA, 10 nmol/L) to the growth medium Dulbecco's Modified Eagle's Medium. From what we have observed, the results show that all of the utilized protocols...
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Stanovení genetické distance výchozích šlechtitelských materiálů řepky ozimé
Papanová, Michaela
Bachelor thesis is compiled from both the literature review and the practical part. The first one contains a treatise on oilseed rape and its main use. Attention is also paid to methods and history of breeding of this crop. An integral part is also an introduction to the concept of genetic distance and the importance of gene banks. The second part contains information about the research focused on determination of genetic distance in selected samples of oilseed rape by ISSR method. The results of the analysis were used to identify possible duplicates in the crop collection as well as to determine the genetic erosion of the samples. Similarly, the results can be used in the breeding itself, especially when selecting the parental components.
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Evaluation of genetic strains polymorphism of \kur{Beauveria bassiana} based on analyses of using microsatellite markers
PUDIL, Jiří
Microsatellites, or Simple Sequence Repeats (SSRs), are highly polymorphic loci present in nuclear and organellar DNA that consist of repeating units of 1-6 base pairs in length. Microsatellites are very effective molecular markers, which allow us to examine various aspects of organisms. The aim of thesis is to carry out analyzes of microsatellites and describe the genetic relationships between individual strains and monosporic isolates, which were derived from these strains. Usind SSR I would like to asses the degree of genetic polymorphism between \kur{Beauveria bassiana} strains, collected during monitoring in the locality of Černá Hora in Šumava. I am also trying to optimize the method of DNA isolation and method of microsatellites analysis. 44 strains of \kur{Beauveria bassiana} isolated from \kur{Ips typographus} were analyzed in total using four primers. Overall, the results that I obtained show high similarity of evaluated strains, and therefore low level of genetic polymorphism. For further work it would be appropriate to enlarge the area, which will be sampled and include sets of strains that were obtained from different developmental stages of the host, as well as other reference strains of entomopathogeníc fungi collected from other localities with similar conditions. For more detailed study it would be useful to use the analysis of ribosomal DNA. Thus selected and evaluated population is likely to provide a more veritable image of strain diversity in observed area.
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Methods of isolation of individual subpopulations of neural stem cells
Zelený, Martin ; Zíková, Martina (advisor) ; Vargová, Ingrid (referee)
Stem cells have huge medical potential, but they are not explored enough for the use in medicine yet. Because of that, their further research is important. One group of the stem cells are neural stem cells, which are differentiating into the neural and glial cells in the brain. Neural stem cells are present at the two areas in the adult brain, subventricular zone and subgranular zone of hippocampus. Neural stem cells are not one homogenous population of cells, but rather various subpopulations with different cell expression. In my bachelor thesis I described the methods, which are used in the research of neural stem cell subpopulations. Described methods focused mainly on the isolation and gene expression. Keywords: neural stem cells, neurogenic zones, molecular markers
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The use of molecular markers for the study of the impact of global change on fungal communities
Faltysová, Julie ; Baldrian, Petr (advisor) ; Koukol, Ondřej (referee)
Fungi are an essential part of life on Earth. Besides their main role in decomposition of organic matter, they belong among key symbiotic organisms and they take part in providing a range of ecosystem processes important for humankind such as primary production and with that related food availability or soil water regulation. For the study of fungal distribution, we often use a method called DNA metabarcoding which combines the use of specific molecular markers, most commonly universal fungal ITS, and high-throughput sequencing. The importance of the method has been rising in the current time of changing environment and it has helped us observe changes in fungal communities due to global change. It seems that primarily various climatic factors such as warming, changes in rainfall patterns and nitrogen deposition have strongly influenced community and fungal guilds composition. Consequently, these changes may potentially have an impact on ecosystem functioning so that their overall balance can be in threat. Key words: molecular markers, DNA metabarcoding, fungal community, warming, climate change, nitrogen deposition
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Differentiation of keratinocytes: molecular markers and potential of influencing them in vitro
Ondrúšková, Denisa ; Filová, Elena (advisor) ; Porubská, Bianka (referee)
Keratinocytes are the most abundant skin cells found in epidermis. They are divided into proliferative basal stem cells, which are in close contact with basement membrane and suprabasal differentiating cells. Basal keratinocytes express K5 and K14 keratins and give rise to differentiating layers via delamination or asymmetric division. The firstly formed layer is stratum spinosum that expresses keratins K1 and K10 and involucrin, and, subsequently, it passes into the stratum granulosum, in which cells express loricrin and profilagrin. The last layer of epidermis is the stratum corneum formed by corneocytes that finally desquamate. Keratinocytes participate in the process of skin regeneration and can be isolated and cultivated. Their cultivation can be affected by various factors, such as selection of suitable materiál (nanofibers/gels) and suitable culture media, which can be enriched with growth factors, platelet lysat, vitamins and other substances. When culturing them, it also depends on whether the cells are entirely immersed in medium or growing on liquid/air interface. To approximate in vivo conditions and to study interaction between cell populations, keratinocytes are often cultured in co-cultures with different cells such as fibroblasts, endothelial cells, monocytes and others....
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Dispersion of freshwater gastropods
Buďová, Jana ; Černý, Martin (advisor) ; Juřičková, Lucie (referee)
Freshwater gastropods (Mollusca: Gastropoda) belong to two tradional taxonomic groups: prosobranchs (Prosobranchia) and pulmonates (Pulmonata). Most of these molluscs are characterized by low vagility. Therefore they usually rely on passive dispersal. Their dispersal vectors are mostly birds and water. Animals can transport snails both externally and internally. Direct methods (capture.mark.recapture, radio - tracking) and genetic methods could be used to study dispersal. For genetic studies can be used many type of molecular markers, but the recently most popular are microsatellites. According to recent studies, the dispersal of freshwater gastropods is probably not as frequent and wide - spread as assumed before. Key words: freshwater snails, dispersal, dispersal vectors, capture - mark - recapture, molecular markers, population genetics
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