|
|
Regulation of bone formation using osteogenic supplements in an osteoporotic model
Krčmářová, Eliška ; Filová, Eva (advisor) ; Zadražil, Zdeněk (referee)
Osteoporosis is a disease of the bone metabolism which is characterised with a decrease of bone substance. The cause of this disease is the imbalance between the creation of a new bone substance by osteoblasts and the resorption of a bone tissue by osteoclasts, in favour of the bone resorption. The risk group of the development of this disease are women after menopause, who naturally register a decline of the estrogen hormone. Estrogen operates as an inhibitor of proosteoclastic factors such as receptor activator of NFκB ligand (RANKL), interleukin (IL)-1, IL-6 or TNF-α. The imbalance of the bone metabolism can also be caused by a disbalance in the production of Prostaglandin E2 (PGE2) and 1α,25-dihydroxyvitamin D3. They are strong mediators which can both stimulate and inhibit an osteoclastogenesis in vitro in concordance with the conditions of the culture/co-culture. This thesis focuses on the examination of an influence of those mediators (PGE2 in the concentration of 10-6 M and 10-8 M; 1α,25-dihydroxyvitamin D3 in the concentration of 10-8 M and 10-9 M) on the osteoclastogenesis from the rat PBMC at the presence of osteoblasts, with or without the combination of proosteoclastic factors macrophage colony-stimulating factor (M-CSF) and RANKL. Osteoclastogenesis was stimulated if PGE2 and...
|
|
|
In vitro cultivation of tapeworm Hymenolepis diminuta - 2
Jandura, Dominik ; Vokřál, Ivan (advisor) ; Raisová Stuchlíková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Dominik Jandura Supervisor: PharmDr. Ivan Vokřál, Ph.D. Title of diploma thesis: In vitro cultivation of tapeworm Hymenolepis diminuta - 2 Aim of this diploma thesis was to obtain cycticercoids of the rat tapeworm (Hymenolepis diminuta), excyst them and find out the conditions for the maximal in vitro incubation period. As the intermediate host mealworm beetle (Tenebrio molitor) infected by the rat feces containing tapeworm eggs was used. Excystment was done using L-cystein and sodium tauroglycocholate. Excysted larvae were cultured in vitro (37 řC, 5 % CO2) in RPMI 1640 medium enriched with other substances chosen according previously published methods. Mainly sheep, mouse or rat liver extracts eventually in combination with yeast extract and sheep bile were used. The effect of tested substances on the cultivation was evaluated by measuring of the tapeworm's growth. The best effect on the grow of the tapeworms was observed using medium containing serum, yeast extract and sheep liver extract where tapeworms achieved length of 1561 µm after 16 days of incubation. The further growth was limited by appearance of pathologic formations.
|
|
|
In vitro cultivation of tapeworm Hymenolepis diminuta - 2
Jandura, Dominik ; Vokřál, Ivan (advisor) ; Raisová Stuchlíková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Dominik Jandura Supervisor: PharmDr. Ivan Vokřál, Ph.D. Title of diploma thesis: In vitro cultivation of tapeworm Hymenolepis diminuta - 2 Aim of this diploma thesis was to obtain cycticercoids of the rat tapeworm (Hymenolepis diminuta), excyst them and find out the conditions for the maximal in vitro incubation period. As the intermediate host mealworm beetle (Tenebrio molitor) infected by the rat feces containing tapeworm eggs was used. Excystment was done using L-cystein and sodium tauroglycocholate. Excysted larvae were cultured in vitro (37 řC, 5 % CO2) in RPMI 1640 medium enriched with other substances chosen according previously published methods. Mainly sheep, mouse or rat liver extracts eventually in combination with yeast extract and sheep bile were used. The effect of tested substances on the cultivation was evaluated by measuring of the tapeworm's growth. The best effect on the grow of the tapeworms was observed using medium containing serum, yeast extract and sheep liver extract where tapeworms achieved length of 1561 µm after 16 days of incubation. The further growth was limited by appearance of pathologic formations.
|
|
|
Somatic embryogenesis of conifers: description of structural development
Kadlecová, Marie ; Albrechtová, Jana (advisor) ; Konrádová, Hana (referee)
Somatic embryogenesis is a very important tool of in vitro cultivation, therefore, it has recently devoted adequate attention. Stages of somatic embryogenesis involve several consecutive steps: induction of embryogenic culture and its proliferation, maturation of embryo, desiccation, germination and subsequent transfer of embryos into young seedlings. All of these steps are directly influenced by many factors, such as a choice of the original explant, time of cultivation, physical conditions or the composition of the culture medium. There are still emerging new studies describing how the culture conditions and, thus, the quality of embryos could be improved. An important prerequisite for further improving the quality of somatic embryos is good knowledge of aculture protocol, i.e. right concentrations of amended substances in the culture medium, the timing of each step, using the suitable explants etc. Another very important thing is good knowledge of development of all anatomical structures established during embryogenesis. Only then it will get closer to the point when somatic embryos will not arise only in the laboratory, but could be successfully used in practice. The object of this work is to summarize existing knowledge about development of all anatomical structures during somatic...
|
|
|
The effects of a hydrogen sulfide donor and garlic derivatives on the meiotic maturation and aging of porcine oocytes
Dvořáková, Markéta ; Sedmíková, Markéta (advisor) ; Miloslava, Miloslava (referee)
Oocyte meiotic maturation is a process of a transformation of the oocyte at the germinal vesicle stage to the oocyte at the stage of second meiotic metaphase. The process called oocyte aging occurs when oocytes are exposed after the completion of meiotic maturation to the prolonged cultivation. This process is characterized by a decrease in oocyte quality. The addition of suitable compounds into the cultivation medium could improve the quality of maturing and aging oocytes. Hydrogen sulfide is a gaseous molecule that is endogenously produced and fulfills signal function in organisms. Alliin and S-allyl cysteine are garlic derivatives that exhibit many positive effects on organisms. Many of them are mediated by their ability of releasing hydrogen sulfide.
We hypothesize that the quality of oocytes in the in vitro cultivation system can be improved by the addition of the hydrogen sulfide donor and garlic derivatives into the cultivation medium. The aim of the study was to evaluate the effects of hydrogen sulfide and garlic derivatives on the meiotic maturation of oocytes, the effects of hydrogen sulfide on oocyte aging and the effects of cultivation with hydrogen sulfide and garlic derivatives on early embryonic development.
It was found out that hydrogen sulfide donor accelerates oocyte nuclear maturation and causes an earlier increase of MPF and MAPK activities. Hydrogen sulfide donor improves activating potential of oocytes. It was found out that endogenous hydrogen sulfide production occurs during aging of procine oocytes and that the hydrogen sulfide production decreases after first 24 hours of prolonged cultivation. The inhibitors of hydrogen sulfide releasing enzymes impaired the quality of aged oocytes. Hydrogen sulfide donor completely suppressed fragmentation of oocytes exposed to prolonged cultivation and improved the quality of aged oocytes treated simultaneously with the inhibitors of hydrogen sulfide releasing enzymes. The presence of hydrogen sulfide donor during the prolonged cultivation of oocytes improved early embryonic development after parthenogenetic activation. It was found out that whereas alliin disrupts meiotic maturation of oocytes, S-allyl cystein has not influenced this process. S-allyl cysteine accelerated the onset of embryo cleavage and reduced ROS levels in maturing and parthenogenetically activated oocytes.
|
|
|
Srovnání asymbiotických kultur in vitro zástupců rodu Cypripedium
Vetter, Martin
The bachelor thesis deals with asymbiotic sowing in vitro in the genus Cypripedium. There are completed information about the genus Cypripedium including the charakteristics of sown species. There is solved composition of culture media for asymbiotic sowing influence of kinetin and 6-benzylaminopurine to germination and influence of added organic constituents such as coconut endosperm, yeast extract and carrot extract. With asymbiotic sowing also binds problems of sterility environment and methodology of surface sterilization of Cypripedium seeds. This treatment i salso important for removing of germination inhibiting substance. The results should contribute to enhancing the methodology of asymbiotic sowing and the influence of organic ingredients used for germination and protocorm development in the studied species of the genus Cypripedium.
|
|
| |
guest ::
