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Modulace intracelulární kaspázové mašinérie s využitím explantátových kultur
Matalová, Eva ; Fleischmannová, Jana ; Norek, Adam ; Míšek, Ivan
Organ explant cultures eliminate animal suffering during experiments but simultaneously allow investigations of intact organ, tissue and cell systems with preserved interactions corresponding to the situation in vivo. Moreover, the explants can be manipulated in several ways using modern micromanipulation techniques (e.g. electroporation, inhibition). We show possible usage of explant culture approaches in embryonal apoptosis research in three models – mouse limb digitalization, tooth germ morphogenesis, separation of middle ear ossicles. Data from specific pharmaceutical inhibitions focused on key apoptotic molecules – caspases - are demonstrated.
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Podíl subtypů P2 receptorů na modulaci glutamatergního přenosu v mozkové kůře
Honzová, Lenka ; Herink, Josef (advisor) ; Hrdina, Radomír (referee)
Glutamate is the major excitatory neurotransmitter in the central nervous system, and there is emerging evidence that its release is subject to presynaptic regulation by P2 receptors. Activation of P2X receptors elicited glutamate release from terminal of dorsal horn neurons of spinal cord and hippocampus whereas activation of P2Y receptors has been shown to inhibit glutamate release in the hippocampus. Glial cells express several subtypes of P2Y receptors and there is growing evidence that these cells are active elements at synapses, they release ATP upon activation of glutamate receptors and participate in the regulation of glutamatergic transmission. The aim of the present study was to investigate the role of P2Y receptors in the regulation of glutamatergic transmission in rat brain cortex; whether uptake of glutamate by glial cells is influenced by P2Y receptor-activation and to identify the subtypes of P2Y receptors involved. Primary cultures of cortical astrocytes obtained from brain hemispheres of newborn rats were used as a model to study the influence of P2Y receptors on the uptake of glutamate. We clarified the role of several P2Y agonists and we identified the subtypes of P2Y receptor involved in the regulation of synaptic concentration of glutamate which may provide the knowledge for...
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The Influence of Elicitation on the Production of Suspension Culture of Trifolium pratense L.
Keřková, Adéla ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Adéla Keřková The Influence of Elicitation on the Production of Suspension Culture of Trifolium pratense L. Summary The paper examined the effect of four concentrations of cadmium chloride on the production of flavonoids and isoflavonoids by the Trifolium pratense L. suspension culture (variety DO-9 and SPRINT). The cultures were cultivated in the Gamborg nutrient media with the addition of 2 mg.l-1 of 2,4-dichlorophenoxyacetic acid and 2 mg.l-1 of 6-benzylaminopurine, at the temperature of 25 o C, 16-hr light/ 8-hr dark period. The elicited and the inspection samples underwent the photometric determination of flavonoids in accordance with the Czech Pharmacopoeia 2005 and the determination of flavonoids via the HPLC metod. The results show that the optimal effect of cadmium chloride on the production of flavonoids and isoflavonoids was manifestated after a 168-hour application of a concentration 0,1µmol (variety DO-9) and after 6-hour application (variety SPRINT).
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Production of secondary metabolites in plant tissue cultures
Provazníková, Věra ; Siatka, Tomáš (advisor) ; Kašparová, Marie (referee)
Věra Provazníková Production of secondary metabolites in plant tissue cultures Experimental work aimed at monitoring of influence of the cerium chlorid as an elicitor on the production of scopoletin in Angelica archangelica L suspension cultures. Cultivation was carried out by on Murashige and Skoog medium in the light and also in the darkness. For the quantification of scopoletin was used technique of high performance chromatography with fluorimetric detection. The findings of the work demonstrated the higher values of the scopoletin amount by the cerium iones elicitation at the cultivation in the dark.
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Explant Culture of Trifolium pratense L.
Šustrová, Lenka ; Kašparová, Marie (advisor) ; Dušek, Jaroslav (referee)
Explant Culture of Trifolium pratense L. A principial precondition for successful elicitation used to increase the production of secondary metabolities is, among other, finding a suitable elicitor, its concentration and the optimal period of time of the action of the elicitor on the plant culture in vitro, which was the aim of the present thesis. The effect was examined of a 6, 24, 48 and 168 hour action of the solution of mercury dichloride (in four concentrations) on the production of flavonoids and isoflavonoids in the callus and suspension culture of Trifolium pratense L. cultivated on a Gamborg medium with an addition of 2 mg.l-1 of 2,4-dichlorophenoxyacetic acid and 2 mg.l-1 of 6-benzylaminopurine, at the temperature of 25řC and 16 hour light/8 hour dark period. The maximal content of flavonoids found by a photometric determination according to Pharmacopoeia Bohemica 2005 was demonstrated in the callus culture (0,636 %) after 6 hour action of the elicitor of the 100 µmol concentration and in the suspension culture (0,492 %) after 168 hour action of the elicitor of the 10 µmol concentration. The maximal content of isoflavonoids found by a HPLC method was demonstrated in the suspension culture after 168 hour action of the elicitor of the 100 µmol concentration (0,03 % of daidzein) and after 168...
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In vitro cultures of medicinal plants - IV.
Kunzová, Lucie ; Tůmová, Lenka (advisor) ; Kašparová, Marie (referee)
In vitro cultures of medicinal plants - IV. Lucie Kunzová Elicitation is one of the methods which can effect the production or accumulation of secondary metabolites in plant cultures. The effect of 6, 12, 24, 48, 72 and 168 hours influence by three concentrations of the abiotic elicitor methylviologen (paraquat) on the rutin production by Fagopyrum esculentum Moench. callus and suspension culture was monitored in this study. The in vitro culture was cultivated on Murashige-Skoog medium with the addition of 10 mg/l of dichlorphenoxyacetic acid (2,4-D) as a growth regulator. The content of rutin was determined by HPLC. The maximal content of rutin in callus culture (0,25 %) was demonstrated after 72 hours of methylviologen elicitation (c1 = 2,1929 · 10-3 mol/l). The maximal content of rutin in suspension culture (0,02 %) was demonstrated after 24 hours of methylviologen elicitation (c3 = 2,1929 · 10-5 mol/l).
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Elicitation of Suspension Culture of Trifolium pratense L.
Sosvorová, Lucie ; Kašparová, Marie (advisor) ; Siatka, Tomáš (referee)
Explant culture of Trifolium pratense L. Summary The objektive of this work was to observe the effect of the elicitors of jasmonic acid and mercuric chloride on the production of flavonoids by the Trifolium pratense L. suspension culture (variety Tempus). By the elicitor of mercuric chloride was observed also its effect to the production of isoflavonoids in the same culture. The cultures were cultivated in Gamborg nutriet media with the addition of 2 mg.l-1 2,4-dichlorphenoxyacetic acid and 2 mg.l-1 6-benzylaminopurine, at the temperature of 25řC and 16-hours light/8-hours dark period. The elicited and control samples underwent the photometric determination of flavonoids in accordance with the Czech Pharmacopoeia 2005 and the determination of isofavonoids via the HPLC method. From the results of the elicitation of Trifolium pratense L. by both elicitors we can see, that more profitable of these is jasmonic acid. Compared to the mercuric chloride there was found a significant incerase of flavonoid production while using a concentration of 5 μmol and operating time 24 hours. Via the HPLC metod were found out the isoflavonoids genistin, genistein and daidzein. We can positively evaluate mainly 168-hours aplication of 10 μmol mercuric chloride, which stimulated the production of all these flavonoids.
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In vitro-cultures of medicinal plants - V
Sedlářová, Marcela ; Tůmová, Lenka (advisor) ; Martin, Jan (referee)
The elicitation of the therapeutically important cultures in vitro is a metod used for increasing the production of secondary metabolites. The effectiveness of elicitation depends on a complex interaction between the elicitor and the plant cell. The elicitation is influenced by many factors: the elicitors specificity, their concentration, the treatment interval, the growth phase of cell culture, the composotion of nutritional medium and the light. A wide array of external stimuli are capable of triggering changes in the plant cell which leads to a cascade of reactions, ultimately resulting in the formation and accumulation of secondary metabolites which helps the plant to overcome the stress factors. This work the influence of UV radiation on the flavonoids production in the callus cultures Genista tinctoria in vitro investigated. The highest production of flavonids was obtained at genistin under the exposure of callus culture to UV radiation of 254 nm wave lenght for the time 300 s with taking of sample after 48 hours. This callus culture grew on SH medium. The content of genistin was 3,03 %. Keywords: elicitation, elicitor, secondary metabolites, UV radiation, Genista tinctoria
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Micropropagation of Lippia dulcis Trev.
Zubíková, Alžběta ; Fernández Cusimamani, Eloy (advisor) ; Michaela, Michaela (referee)
Lippia dulcis Trev. (Verbenacea) is a perennial herb with a creeping stem from Central America. It is known for its healing properties for problems with the respiratory or digestive system, but also its sweet taste, which causes terpene hernandulcin because of which the leaves of Lippia dulcis Trev. are 1,000 times sweeter than sugar. Generatively, the plant reproduces by seeds and vegetatively by cuttings.
The main objective of this study was to investigate the influence of cytokinins kinetin (KIN) and 6-benzylaminopurine (BAP) with the addition of auxin indole-3-acetic acid (IAA) on the growth and regeneration of plants from nodal segments Lippia dulcis Trev. in vitro conditions.
Nodal segments were cultured on 6-variants of MS medium supplemented with growth regulators at concentrations of 0.5; 0.75 and 1.0 mg / l BAP and KIN with combination 0.5 mg / l IAA and one variant of pure MS medium as a control. Nodal explants were cultured for 35 days (the time of subculturing) and measured every 7 days. The following parameters were evaluated: regeneration, number of shoots, length of shoots, number of nodes, number and length of roots.
The measurement results show that a combination of growth regulators auxin-cytokinin have no positive influence on the growth and regeneration Lippia dulcis Trev. in vitro conditions. The best regeneration (100%), growth (on an average length of the shoots 3.05 +- 1.42) and the highest number of newly created points (5.79 +- 2.4) occurred at the explants cultured on a clean MS medium without addition of growth regulators. Among the variant with pure MS and media variants with the addition of growth regulators are statistically significat defferences, but without a favorable influence on the development of nodal explants.
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Explant cultures of Higher plants 29
Faltýnková, Jana ; Dušková, Jiřina (advisor) ; Tůmová, Lenka (referee)
Faltýnková, J.: Explant cultures of higher plants 29. Diploma thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, 2008 The goal ofmythesis was to verify biotransformation posibility of theDatura meteloides culture. Hydroquine - the precursor of arbutin was added into these cultures. Precursor used in this study was prepared in concentration of 100 mg/l and 200 mg/l. Exposition of precursor was 24, 48, 72, 96, 120, 144 and 168 hours under constant conditions. By the qualitative and quantitative analysis I found the presence of studyied arbutin metabolite in all observed samples. The biggest arbutin volume was obtained after 168 hours of cultivation with precursor of concentration at 200 mg/l ( 3,05%). The in_uence of precursor's concentration and exposition time to the _nal product was thereby con_rmed.
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