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The Role of Cytoskeleton and Phospholipids in signaling During Plant Defense Response
Angelini, Jindřiška ; Dušková, Jiřina (advisor) ; Jahodář, Luděk (referee) ; Grančai, Daniel (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Department of pharmaceutical botany and ecology Candidate PharmDr. Jindřiška Matoušková Supervisor Ass. Prof. RNDr. Jiřina Dušková, Csc Title of Doctoral Thesis The role of cytoskeleton and phospholipids in signaling during plant defense response Upon abiotic and biotic cues, plant surveillance system simultaneously triggers a variety of signaling events to develop an effective defense response within a few hours. On the molecular level, signaling period is the chain of many consecutive and fast biochemical reactions. Tracking of one particular signaling event e.g. during plant-pathogen interactions is quite difficult task. Presented work is focused on several components of this intricate defense network; namely on the role of salicylic acid, phospholipid signaling and cytoskeleton dynamics and their mutual interaction coupled with downstream changes of plant metabolom. Live imaging can help to elucidate localization and dynamics of particular, fluorescently labeled proteins during defense response. Plenty of important events occurring in plant- -pathogen interactions take place in plasma membrane: where pathogen perception, exocytosis, endocytosis occure and ion channels and carriers are located. Also phosphatidic...
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The Evaluation of Zearalenone Pressencence in Food and Feed in the Czech Republic
Čtvrtečková, Tereza ; Karlíčková, Jana (advisor) ; Jahodář, Luděk (referee)
1 ABSTRACT ČTVRTEČKOVÁ T.: The Evaluation of Zearalenone Pressencence in Food and Feed in the Czech Republic. Hradec Králové 2013, 92 pages. Doctoral thesis at the Department of Pharmaceutical Botany and Ecology, Faculty of Pharmacy in Hradec Králové, Charles University in Prague. Supervisor: PharmDr. Jana Karlíčková, Ph.D. Zearalenone, a mycotoxin characteristic for its estrogenic effects, is produced by certain species of Fusarium genus. It appears in cereal grains, especially in corn. The thesis presents an overview of contemporary findings regarding mycotoxins of the zearalenone type in the territory of the Czech Republic. The focus lies on current assessment of zearalenone presence in food and feed. A summary of latest information on the legislative regulations of zearalenone in food, feed and animal products is provided, including zerealenone analysis methods, their characteristics and trends. The thesis also draws attention to the importance of mycotoxin occurence prevention.
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Ecotoxicological screening of the medicaments - Paralen tbl. and Panadol tbl.
Janečková, Petra ; Vytlačilová, Jitka (advisor) ; Jahodář, Luděk (referee)
(EN) Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Petra Janečková Consultant: Mgr. Jitka Vytlačilová Title of thesis/dissertation: Ecotoxicological screening of the medicaments - Paralen tbl. and Panadol tbl. Paracetamol is an often-used medicinal substance from the group of analgetics-antipyretics. This thesis evaluates the influence of paracetamol contained in pharmaceuticals Paralen 500 tbl. and Panadol tbl., and of the paracetamol analytical standard on the environment. Three ecotoxicological tests and representatives of three trophic-level organisms were selected: a multi-generational test with the protozoan Tetrahymena pyriformis Ehr., a rapid screening test RapidtoxkitTM with the crustacean Thamnocephalus platyurus Packard and an inhibition test of the germination of Sinapis alba L. seeds. The highest tested concentration of paracetamol was 2341,67 mg/l. The tests evaluated the inhibition of the protozoan's growth; the inhibition of the crustacean's food intake, and the inhibition of seed germination in higher plants. The most sensitive organism was the producer of Sinapis alba L., the least sensitive was the consumer Thamnocephalus platyurus Packard. The values of IC50 were calculated based on the results of...
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Determination of nitrates in herbal products and evaluation of their antimicrobial activities
Drdová, Diana ; Vytlačilová, Jitka (advisor) ; Jahodář, Luděk (referee)
Determination of nitrates in herbal products and evaluation of their antimicrobial activities (Diploma thesis) Diana Drdová Charles University in Prague Faculty of Pharmacy in Hradec Kralove Department of Pharmaceutical Botany and Ecology The aim of this diploma thesis is to ascertain the content of nitrates in herbal teas and evaluate the microbiological toxicities. We tested 47 specimens of herbal teas, bought in Pharmacies and in herbalist shops. Occurrence of moulds and yeasts in specimens was measured with a technique of counting colonies, cultivated at 25 žC on agar plates with yeast extract, glucose and chloramphenicols after five days of incubation. Occurrence of Escherichia coli in specimens was measured with another technique of counting colonies cultivated at 44 žC on agar plates with 5-Bromo-4-chloro-3-indolyl-_-D-Glucoronid, after 24 hours of incubation. The presence of nitrates was measured with a method of High Performance Liquid Chromatography with a detection of UV light spectrum, in accordance with standards setting limits for nitrates in food. Nearly each specimen contained moulds, of which mostly Rhizopus niger (47.37 %) and Aspergillus niger (33,33 %). No yeasts were found in tested specimens. The Gram-negative bacteria Escherichia coli was found only in one herb tea - Milk...
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Biological Activity of Cichorium Intybus Compounds
Málková, Marta ; Jahodář, Luděk (advisor) ; Karlíčková, Jana (referee)
Marta Málková: Biologická aktivita obsahových látek Cichorium intybus. Rigorózní práce, Univerzita Karlova v Praze, Farmaceutická fakulta v Hradci Králové, Katedra farmaceutické botaniky a ekologie, Hradec Králové 2010, s.57. Klíčová slova : Cichorium intybus, čekanka obecná, arnikolid Biologická aktivita byla stanovována u extraktů získaných z plodů a kořenů Cichorium intybus. Použit byl ethanolový, vodný a dichlormetanový extrakt ze sušených kořenů a suchý vodný extrakt z plodů a z něj získaná frakce obsahující fenolické kyseliny. Významný inhibiční efekt na acetylcholinesterázu a butylcholinesterázu nebyl zaznamenán u žádného z extraktů. Nejvyšší obsah fenolických látek a zároveň nejvyšší antioxidační aktivita byla zjištěna u frakce obsahující fenolické kyseliny. Dále byly všechny extrakty hodnoceny pomocí tenkovrstvé chromatografie, která prokázala přítomnost sitosterolu v dichlormethanovém extraktu z kořenů Cichorium intybus. Na základě výsledků TLC byla pomocí HPLC v tomto extraktu zjištěna přítomnost laktonu arnikolidu. ABSTRACT Marta Málková: Biological activity of Cichorium intybus compounds. Rigorous thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany and Ecology. Hradec Králové, 2010, s.57. Keywords : Cichorium intybus,...
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Usage of the annelid Tubifex tubifex Müll. as a model organism for biological activity assessment
Vytlačilová, Jitka ; Jahodář, Luděk (advisor) ; Kočí, Vladimír (referee) ; Fiala, Zdeněk (referee)
The aim of the presented doctoral thesis was to use the sludge worm Tubifex tubifex as a model for biological activity testing. In the presented study, the Tubifex worm was used to evaluate photosensibilisation, i.e. sensibilisation of an organism towards UVA radiation (365 nm; radiant flux density of 350 μW/cm2) after contact with certain substances. The tested substances included proven photosensibilizing agents - rose bengal, khellin and xanthotoxin; isolated natural substances - thiophene polyacetylene umbelliferon, scopoline, and scopoletine; synthetic compounds - salicylanilides and thiosalicylanilides; and penicillin drugs Augmentin and Ospamox and their active substance standard amoxicillin. The final evaluation of photosensibilizing properties of the substances was carried out according to the Commission Directive 2000/33/EC PHOTOTOXICITY - IN VITRO 3T3 NRU PHOTOTOXICITY TEST. The test was performed in 24-well plates where one set of plates with the photosensibilizing agent was irradiated and the second set was placed in an incubator (20±2řC) as a dark control without any light presence for the whole period of the experiment. The plates were evaluated immediately after the given period of irradiation (15, 30, 45, 60, 120, and 240 minutes). Mortality and percentual number of damaged individuals were...
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Biologically active metabolites of plants. 10. Alkaloids of Fumaria officinalis L. and their biological activity
Šafratová, Marcela ; Cahlíková, Lucie (advisor) ; Jahodář, Luděk (referee)
Šafratová M.: Biologically active metabolites of plants 10. Alkaloids of Fumaria officinalis L. and their biological aktivity. Charles University in Prague, Faculty of Pharmacy in Hradec Králové. Department of Pharmaceutical Botany and Ecology, Hradec Králové, 2013. Diploma thesis. 70 s. The object of this diploma thesis was to process the pool of alkaloids from Fumaria officinalis L. (Fumariaceae). Using column chromatography and preparative TLC three alkaloids were isolated which have been subjected to structural analysis (MS, NMR). The first substance was identified as isoquinoline alkaloid stylopin, others isolated alkaloid was sinactin and third isolated alkaloid was fumarilin. All three alkaloids have been previously isolated from F. officinalis. For all three alkaloids were tested for their inhibitory activity against human erythrocyte acetylcholinesterase (HuAChE) and human serum butyrylcholinesterase (HuBuChE). For stylopine IC50 values were determined IC50 HuAChE ˂ 1000 µM and IC50 HuBuChE ˂ 1000 µM. For sinactine were measured IC50 HuAChE = 632 ± 22 µM and IC50 HuBuChE ˂ 1000 µM. Values for fumariline IC50 HuAChE ˂ 1000 µM and IC50 HuBuChE = 330 ± 12 µM. In comparison with standards were stylopine, sinactine and fumariline not very active. None of the isolated compounds are therefore...
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Determination of artemisinin in Artemisia annua
Špundová, Jana ; Jahodář, Luděk (advisor) ; Karlíčková, Jana (referee)
This studies describes the determination of artemisinin in Artemisia annua L. cultivated in Czech Republic in 2006. Plant was harvested in four vegetative stages (intensive vegetative growth stage, bud formation, cease bloom stage and maturity seed stage). Plant was shade dried on the air and extracted with an organic solvent hexane for 17 hours and then filtrated. Artemisinin was confirmed by thin layer chromatography with artemisinin standard use. The concentration of artemisinin was determinated by reversed phase high performance liquid chromatography with constant constitution of mobil phase and UV detection with wave length 295 nm. Maximum artemisinin concentration occured at the bud formation stage (0,030%).
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Pharmacognostic study of Alnus spp.
Chlebek, Jakub ; Jahodář, Luděk (advisor) ; Karlíčková, Jana (referee)
Charles Univerzity in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Degree work term: Pharrnacognostic study of Alnus spp. Student: Jakub Chlebek Abstract The degree work Pharmacognostic study of radix Alnus spp. consists of theoretical and experimental part. Theoretical part contains botanical characteristic of the three home species of Czech Republic (Alnus glutinosa, Alnus incana, Alnus rugosa), phytochemical review of the compounds of the genus Alnus and their biological acivities. Experimental part deals with phytochemical and biological study of the roots of Alnus glutinosa. Dried roots of the black alder (Alnus glutinosa) were extracted with Me2CO-H2O (7:3). Final extract was chromatographed over test silica gel colum using a MeOH- CHCl3 gradient (0, 1, 2, 4, 8, 16, 32, 100 %). Elution gave 36 fractions. All the separations were analysed by TLC (hexane-EtOAc MeOH, 2:2:1). The sterols were detected in the fraction No. 7 by TLC, melting point and GC-MS. Major present sterols were β-sitosterol, β-sitostanol, minor sterols campestanol, campesterol. Fractions No. 10 and 19 were analysed by HPLC. Antioxidant activity of the fraction No. 19 was measured with DPPH radical and was low (EC50 0.32 mg/mL). Test silica gel colum experience were used in analysis on...
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