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Preparation of STAT3-knockout glioblastoma multiforme cell line using CRISPR-Cas9 editing
Papinčáková, Dominika ; Skálová, Lenka (advisor) ; Laštovičková, Lenka (referee)
Charles University Fakulty of Pharmacy in Hradec Králove Department of Biochemical Sciences Candidate: Bc. Dominika Papinčáková Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: Preparation of STAT3-knockout glioblastoma multiforme cell line using CRISPR-Cas9 editing Glioblastoma multiforme (GBM) is one of the most common primary brain tumours in adults and one of the most fatal cancers in humans A key oncogene in GBM development and progression is the signal transducer and activator of transcription 3 (STAT3). It is one of the mediators of tumorigenesis, plays a strong pro-survival role in GBM, and is permanently activated in GBM, leading to increased cell proliferation, invasiveness, and angiogenesis. The aim of this work was to establish a GBM STAT3-knockout (STAT3-KO) cell line. The U87MG cell line and the CRISPR-Cas9 gene knockout method were used for this purpose. In a first step, we investigated the possibility of silencing STAT3 expression by siRNA and the suitability of this cell type to pinpoint the mechanism for CRISPR gene editing into the cell. In the second step, we transfected the cells using the STAT3 gene knockout kit (CRISPR) and tested the knockout success. In the established STAT3-KO GBM cell line, we confirmed STAT3-KO at the DNA, RNA and protein levels. We...
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Preparation of STAT3-knockout glioblastoma multiforme cell line using CRISPR-Cas9 editing
Papinčáková, Dominika ; Skálová, Lenka (advisor) ; Laštovičková, Lenka (referee)
Charles University Fakulty of Pharmacy in Hradec Králove Department of Biochemical Sciences Candidate: Bc. Dominika Papinčáková Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: Preparation of STAT3-knockout glioblastoma multiforme cell line using CRISPR-Cas9 editing Glioblastoma multiforme (GBM) is one of the most common primary brain tumours in adults and one of the most fatal cancers in humans A key oncogene in GBM development and progression is the signal transducer and activator of transcription 3 (STAT3). It is one of the mediators of tumorigenesis, plays a strong pro-survival role in GBM, and is permanently activated in GBM, leading to increased cell proliferation, invasiveness, and angiogenesis. The aim of this work was to establish a GBM STAT3-knockout (STAT3-KO) cell line. The U87MG cell line and the CRISPR-Cas9 gene knockout method were used for this purpose. In a first step, we investigated the possibility of silencing STAT3 expression by siRNA and the suitability of this cell type to pinpoint the mechanism for CRISPR gene editing into the cell. In the second step, we transfected the cells using the STAT3 gene knockout kit (CRISPR) and tested the knockout success. In the established STAT3-KO GBM cell line, we confirmed STAT3-KO at the DNA, RNA and protein levels. We...
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Determination of ovarian function parameters
Jará, Barbora ; Skálová, Lenka (advisor) ; Laštovičková, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Thesis title: Determination of ovarian function parameters Supervisor: prof. RNDr. Lenka Skálová, Ph. D. Consultant: RNDr. Michala Vosátková, MBA Created by: Barbora Jará The levels of hormones that control of ovaria are important markers of development and health status of the female organism. Laboratory examination and correct interpretation of the results are important for proper diagnostic procedures. Among the hormones of ovarian function, the level of estradiol, progesterone, luteinizing and follicle-stimulating hormones is most often tested. Separation and immunochemical methods are used. For common clinical practice, immunochemical methods are particularly suitable due to their availability, relative accuracy, unpretentiousness, and speed of processing. The impossibility of standardizing individual methodological approaches represents the shortcoming of these methods. A series of examinations and interpretation of results can be performed using immunochemical methods only within one workplace. Keywords: estradiol, immunochemical methods, ECLIA, CMIA, RIA
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Liability for damage under civil law and under commercial law
Laštovičková, Lola ; Švestka, Jiří (advisor) ; Salač, Josef (referee)
The purpose of my thesis is to compare the conformity and diversity between legal provisions of liability for damages in czech civil and commercial law. The thesis is divided into four chapters and its core lies in Chapter Three. First chapter subdivided into three parts deals with characteristics of liability for damages in general, further contains the location of this liability among other types of liability and the last part concerns with the functions of it. Chapter Two is concentrated on the Principles of European Tort Law. Part One describes the europeisation of tort law, which is mostly connected with spontaneous projects of soft law. In Part Two there is reffered to general conception of the Principles and some particular provisions in detail. I have endeavoured to draw a comparison between almost every field of liability for damages in czech civil and commercial law and these Principles. As I have mentioned the main part of my thesis rests in Chapter Three. The chapter consists of eight parts. At first I have explained the relationship between Czech Civil Code and Commercial Code regarding liability for damages. Legal provisions in both codes are sometimes duplicate, on the other hand in some parts different but often without any relevant reason. Subsequent part highlights the importance...
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Influence of albendazole on the antioxidant system of alfalfa in real conditions
Lisoňková, Monika ; Szotáková, Barbora (advisor) ; Laštovičková, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Monika Lisoňková Supervisor: prof. Ing. Barbora Szotáková, PhD. Title of diploma thesis: The effect of albendazole on the antioxidant system of alfalfa in real conditions Anthelmintics are drugs used to treat diseases caused by parasitics worms. In the Czech republic they are mainly used in veterinary medicine. Benzimidazole anthelmintic albendazole is used in cattle and sheep, during its biotransformation metabolites are formed, which are excreted together with the parent compound in faeces. Application of faeces from treated cattle to the field may affect non-target organisms, occurring in the field. An example of such organisms are agricultural crops, including alfalfa (Medicago sativa). Plants do not have the ability to eliminate the absorbed drugs and its metabolites, therefore they are deposited in plants and can affect the antioxidant system of plants. The aim of this study was to monitor the effect of albendazole on the antioxidant system of alfalfa (M. sativa) in real conditions. Plants were affected by faeces of sheep treated with albendazole. Subcellular fractions were prepared from harvested plants in the years 2019 and 2020. The concentrations of photosynthetic pigments...
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Liability for damage under civil law and under commercial law
Laštovičková, Lola ; Švestka, Jiří (advisor) ; Salač, Josef (referee)
The purpose of my thesis is to compare the conformity and diversity between legal provisions of liability for damages in czech civil and commercial law. The thesis is divided into four chapters and its core lies in Chapter Three. First chapter subdivided into three parts deals with characteristics of liability for damages in general, further contains the location of this liability among other types of liability and the last part concerns with the functions of it. Chapter Two is concentrated on the Principles of European Tort Law. Part One describes the europeisation of tort law, which is mostly connected with spontaneous projects of soft law. In Part Two there is reffered to general conception of the Principles and some particular provisions in detail. I have endeavoured to draw a comparison between almost every field of liability for damages in czech civil and commercial law and these Principles. As I have mentioned the main part of my thesis rests in Chapter Three. The chapter consists of eight parts. At first I have explained the relationship between Czech Civil Code and Commercial Code regarding liability for damages. Legal provisions in both codes are sometimes duplicate, on the other hand in some parts different but often without any relevant reason. Subsequent part highlights the importance...
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Loyalty Program for Retail Chain Tesco – Clubcard
Laštovičková, Lucie ; Postler, Milan (advisor) ; Szabo, Peter (referee)
This thesis deals with loyalty program of retail chain Tesco -- Clubcard loyalty program. The thesis is divided into two parts. The first part is focused on terms explanation -- e.g. marketing and commercial communication, sales promotion and it's techniques and also loyalty and loyalty programs itself. The beginning of practical part firstly describes current trends on the retail market. The following competitive analysis of retail chains is focused on loyalty programs. Next part deals with Tesco company and it's Clubcard loyalty program. The main contribution is represented by marketing research focused on Tesco customers's opinion about Clubcard program.
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Private labels and how they are perceived by consumers
Laštovičková, Lucie ; Zamazalová, Marcela (advisor) ; Mottlová, Libuše (referee)
Private label products encompass all merchandise sold under a retailer's brand. This brand can be the retailer's own name or a name created exclusively by that retailer. Private labels definitely mean for all three concerned parties (producer, retailer and consumer) many advantages, but also some disadvantages.We can divide them into three categories: economy, standard and premium labels. The most developed private label country in Europe is Switzerland. The private label share reaches about 45%. The share of private labels in Czech republic is almost 20%. At present almost all business chains have their own private label. Private labels are most widespread in discount shops The result of my questionnaire research, says: 67% of informants buy private labels regularly. The greatest advantage for consumers is a low price. On the other hand, the disadvantage is their lower quality and unattractive cover.
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