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Naftochinony - substancje barwne i alkaloidy pirolizydynowe były badane w tkance kalusowej i kulturze zawiesinowej Arnebia euchroma. W celu rozdzielenia frakcji oraz przeprowadzenia analizy jakościowej i ilościowej zastosowano metody chromatografii kolumnowej (CC), chromatografii cienkowarstwowej (TLC) oraz wysokociśnieniowej chromatografii cieczowej (HPLC). Najwyższą zawartość szikoniny (1850 mg 1-1) znaleziono w kulturze zawiesinowej hodowanej w pożywce Davydenkov Arnebia (DA) uzupełnionej jasmonianem metylu - MJ (100 uM). W kontroli (pożywka DA bez MJ) wykryto tylko 280 mg l-1 szikoniny. W tkance kalusowej hodowanej na pożywce DA z pikloramem (10 mg l-1) oznaczono acetyloszikoninę i ślady alkaloidów pirolizydy- nowych (PA). Pikloram zwiększał wskaźnik wzrostu kalusa lecz jednocześnie zmieniało się zabarwienie tkanki z czerwonej na żółto-zieloną. Zarówno kalus jak i kultura zawiesinowa miały 11 lat. Tkanki te przez cały okres były hodowane na pożywce Davydenkova, która w powyższej pracy została użyta do kontrolnych badań porównawczych.
Extracts of Rhodiola Kirilowii plant roots (several years old plant) were studied by HPLC methods. The aim of these studies was searching for active substances: salidroside, tyrosol, rosavin, cinnamyl alcohol and triandrine. In studied extracts mentioned above substances were not found. On chromatograms only one peak (Rt 8,9 min., maximal absorbance at X =220 nm) of notidentified substance was observed. Further investigations are in progress.
An efficient plant micropropagation system was established using shoot tips and stem segments of Lithospermum canescens and Onosma paniculatum (Boraginaceae). A high frequency of axillary shoot formation was achieved on LS [15] medium with 0.5 mg/1 kinetin and 2.0 mg/1 BAP (average shoot formation per explant was 6.18 ± 1.89 for L. canescens and 3.01 ± 0.82 for 0. paniculatum). The best results were obtained with 1x8 clone of L canescens. Axillary branching and root formation were the highest (8.22 shoot formation, 46.7% of rooting). Contamination of the explants with Rhodococcus fascians D188 increased the number of newly formed shoots of 0. paniculatum to 4.6 ± 0.72, while the propagation process of L. canescens was not affected. The highest percentage of rooted shoots of L canescens was observed on V, B5 medium [14] with 0.2 mg/1 1BA (from 27% to 46.7%, depending on plant line), while in the case of regenerated shoots of 0. paniculatum hormone-free LS medium induced roots more effectively (36.7%) than V2 B5 medium with 0.2 mg/1 1BA (25%).
The role of sodium nitroprusside (SNP, 10 lM), a nitric oxide (NO) donor, and/or methyl jasmonate (MJ, 100 lM) spiked with L-phenylalanine (PHEN, 100 lM) and additional sucrose (S; 30 g l-1), in taxane production and phenyl ammonia lyase (PAL) activity in cultures of two Taxus media x var. Hicksii transgenic root lines (ATMA and ATM) carrying the taxadiene synthase transgene was investigated. SNP addition, when applied together with MJ and/or PHEN, resulted in paclitaxel production only in ATMA cultures. The application of the NO donor gave the highest paclitaxel content (7.56 mg l-1) in the combination of SNP+S+MJ+PHEN, after 2 weeks of treatment in the ATMA root line. In ATM cultures, taxane production was not affected by SNP. In both ATMA and ATM lines the highest total (intra+extracellular) paclitaxel yield was determined when elicited with MJ+PHEN, and amounted to 10.78 mg l-1 at 1 week and 1.63 mg l-1 at 2 weeks of treatment, in cultures of ATMA and ATM lines, respectively. The excretion of paclitaxel was observed only in ATMA cultures, with the highest level (2.34 mg l-1) obtained after elicitation with S+MJ+PHEN. The comparison of PAL activity in the two root lines revealed that this enzyme was almost 3-times more active in ATM than ATMA roots. An increase in both PAL activity and paclitaxel production was only observed in ATMA cultures growing in medium supplemented with S+MJ+PHEN.
Antibacterial and antifungal activities of Lithospermum ccinescens roots extract and acetylshikonin, isobutyrylshikonin — naphthoquinone derivatives isolated from them were investigated by measuring their minimal inhibitory concentrations (MIC). L. canescens extract was found to have strong antibacterial and antifungal activities against investigated Gram-positive bacteria: Staphylococcus aureus 1251, Enterococcus faecalis ATCC 8040, and yeast-like fungi strain Candida albicans PCM 1409 PZH and comparable to chloramphenicol and amphotericin B activities. However, the extract potency against Gram-negative bacteria such as Escherichia coli PZH 026 B6 and Pseudomonas aeruginosa S. 85/2 was several times lower. Acetylshikonin and isobutyrylshikonin showed strong anti­biotic activity. The effect of these compounds on the S. aureus FDA 209P was similar as effect of chloramphenicol.
In this study, an efficient method to enhance phenolic compound production in the in vitro cultured shoots of Polyscias filicifolia was developed. The phenolic compound content in P. filicifolia has not yet been reported. Shoots were treated with methyl jasmonate (JM) or salicylic acid (SA) at doses of 50, 100, or 200 μM. HPLC-UV-VIS and LC-MS techniques were used for the determination of chlorogenic, caffeic, and ferulic acids. The total phenolics and flavonoids were quantified, and the antioxidant capacity of plant extracts was determined using DPPH and ABTS methods. Finally, the cytotoxic activity of P. filicifolia extracts in normal (HaCaT) and cancer (A549) cells was investigated. Further, the effect of the extracts on cisplatin cytotoxicity was assessed. The elicitors significantly enhanced phenolic production compared to that in untreated shoots and leaves of intact plants. Chlorogenic acid was the most abundant compound with the highest yield of 5.03 ±0.25 mg/g DW after treatment with 50 μM SA. The total flavonoid and phenolic content was significantly and dose-dependently influenced by JM. The highest antioxidant capacity was noted in extracts derived from shoots grown on media supplemented with 50 μM SA and 200 μM JM; these doses were used for further cytotoxic activity investigations. The extracts from JM or SA treatments reduced cancer cell viability and increased their mortality, whereas the extract from JM treatment exhibited protective effect on normal cells. Moreover, the comparison of cytotoxic properties of plant extracts and cisplatin indicated that plant phenolic compounds in combination with anticancer drugs could reduce the detrimental effect of the latter on human cells.
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