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Use of polymerase chain reaction (PCR) technique for identification of probiotic microorganisms in milk diary product
Horká, Markéta ; Fialová, Lenka (referee) ; Smetana, Jan (advisor)
In this bachelor thesis, probiotic bacteria and their beneficial effects on human health were described. Several methods that are used to identify microorganisms were also stated. At the beginning of the experimental part, DNA was isolated from a selected dairy product. DNA extraction was made in two ways, both of which provided sufficiently high-quality DNA. The prepared DNA was subjected to polymerase chain reaction using specific primers. The presence of bacteria of the genus Lactobacillus, Bifidobacterium and the species Lactobacillus acidophilus, which were declared by the yogurt manufacturer, was proved by electrophoresis on agarose gel.
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The study of interaction of platinum cytostatics with BCRP drug transporter using MTT cytotoxicity test
Horká, Markéta ; Čečková, Martina (advisor) ; Štaud, František (referee)
AAbbssttrraacctt The resistance of oncological diseases to cytostatic treatment is considered as one of the most serious problems of cancer treatment. One of the mechanisms by which cancers develop resistance is the overexpression of efflux transport proteins that limit intracellular concentrations of substrate agents. This work is focused on study of the interaction between clinically used platinum cytostatics (cisplatinum, carboplatinum and oxaliplatinum) and the efflux transport protein - BCRP. The experiments were made on MDKCII (parental cell line) and MDCKII-BCRP (cells were transfected by the gene of BCRP) cell lines. The results of the experiment with oxaliplatinum did not confirm any effect of BCRP on viability of BCRP transfected cells. Cisplatinum and carboplatinum tests provided evidence that BCRP transporter plays an important role in resistance development. The cells that were transfected by the gene of BCRP showed remarkably enhanced viability than parental MDCKII cells. However, the inhibition of BCRP transporter by specific inhibitor fumitremorgin C did not affect the viability of MDCK-BCRP cells. Such results suggest that the resistance of transfected cells was developed by other mechanism. Enhanced resistance could be caused by EGFP which is used as a reporting protein in preparation of...
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The study of interaction of platinum cytostatics with BCRP drug transporter using MTT cytotoxicity test
Horká, Markéta ; Čečková, Martina (advisor) ; Štaud, František (referee)
AAbbssttrraacctt The resistance of oncological diseases to cytostatic treatment is considered as one of the most serious problems of cancer treatment. One of the mechanisms by which cancers develop resistance is the overexpression of efflux transport proteins that limit intracellular concentrations of substrate agents. This work is focused on study of the interaction between clinically used platinum cytostatics (cisplatinum, carboplatinum and oxaliplatinum) and the efflux transport protein - BCRP. The experiments were made on MDKCII (parental cell line) and MDCKII-BCRP (cells were transfected by the gene of BCRP) cell lines. The results of the experiment with oxaliplatinum did not confirm any effect of BCRP on viability of BCRP transfected cells. Cisplatinum and carboplatinum tests provided evidence that BCRP transporter plays an important role in resistance development. The cells that were transfected by the gene of BCRP showed remarkably enhanced viability than parental MDCKII cells. However, the inhibition of BCRP transporter by specific inhibitor fumitremorgin C did not affect the viability of MDCK-BCRP cells. Such results suggest that the resistance of transfected cells was developed by other mechanism. Enhanced resistance could be caused by EGFP which is used as a reporting protein in preparation of...
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