Isolation of three dibenzocyclooctadiene lignans from in vitro cultures of Schisandra chinensis (Turcz.) Baill.--the first report.

Three dibenzocyclooctadiene lignans: deoxyschizandrin (1), gomisin A (2) and schizandrin (3) were isolated from biomass extracts of Schisandra chinensis (Turcz.) Baill. shoot-differentiating callus cultures. The mentioned lignans were not isolated earlier from in vitro cultures of this plant species. This is the first report concerning on isolation of dibenzocyclooctadiene lignans from in vitro cultures of Schisandra chinensis.

Lignans in Schisandra chinensis in vitro cultures.

Contents of schisandrol A and schisandrol B were determined in methanolic extracts of biomass from in vitro cultures of Schisandra chinensis (Turcz.) Baill. (Schisandraceae) using an HPLC method. The biomass was cultured on six variants of Murashige and Skoog (MS) medium containing different concentrations of growth regulators, cytokinin (BAP) and auxin (NAA). The contents of both lignans were different and dependent on the MS medium variant. The maximal contents of schisandrol A (70.54 mg/100 g d.w.) and schisandrol B (86.41 mg/100 g d.w.) were considerable in comparison with plant material: leaves (29.69 and 34.50 mg/100 g d.w.) and fruits (132.39 and 109.40 mg/100 g d.w., respectively). This is the first report on the quantitative analysis of schisandrol A and schisandrol B in Schisandra chinensis in vitro cultures.

Free phenolic acids in Ruta graveolens L. in vitro culture.

Eight phenolic acids were determined using HPLC method in methanolic extracts of Ruta graveolens L. (Rutaceae) shoots cultured in vitro on four variants of Linsmaier and Skoog (LS) medium differing in contents of growth regulators, NAA and BAP (ranging between 0.1-3.0 mg/l). Four compounds: protocatechuic, vanillic, syringic and p-coumaric acid were detected and quantified. The total content of metabolites was dependent on LS medium variants. The contents of protocatechuic acid, quantitatively dominating on all tested LS medium variants, were considerable (from 67.15 to 93.24 mg/100 g d.w.) in comparison with its contents in the plant material under analysis (46.36 to 218.27 mg/ 100 g d.w.). This is the first report of the isolation of protocatechuic acid from an in vitro plant culture.

Indole compounds in fruiting bodies of some selected Macromycetes species and in their mycelia cultured in vitro.

Methanol extracts of fruiting bodies of two purportedly edible species of higher fungi (Macromycetes)- Tricholoma equestre (L.: Fr.) Kumm. and Xerocomus badius (Fr.) Kühn.ex Gilb., collected from a natural environment and of biomass obtained from in vitro cultures were assayed for contents of nine indole compounds. Common indole compounds in extracts from fruiting bodies of both species were: tryptophan, tryptamine and serotonin, in mycelial extracts from in vitro cultures: tryptophan and tryptamine. Contents of the compounds investigated varied widely, from 0.182 to 2.851 mg/100g d.w. in fruiting bodies and from 0.322 to 2.096 mg/100g d.w. in biomass from in vitro cultures, respectively.

Accumulation of biologically active furanocoumarins in agitated cultures of Ruta graveolens L. and Ruta graveolens ssp. divaricata (Tenore) Gams.

This study was designed to investigate the dynamics of accumulation of linear furanocoumarins (psoralen, bergapten, xanthotoxin, isopimpinellin, imperatorin) and their biogenetic precursor, umbelliferone, in agitated cultures of Ruta graveolens L. and Ruta graveolens ssp. divaricata (Tenore) Gams during 6-week growth cycles. The metabolites under study were almost exclusively accumulated in the cultured biomass. The total content of all metabolites increased 4.8- and 2.0-fold, in R. graveolens and R. graveolens ssp. divaricata cultures, respectively. Xanthotoxin and bergapten, which are the most important therapeutic compounds, were the dominating metabolites in cultures of both plants. The maximum content of xanthotoxin (25.0 mg/100 g dry wt.) and bergapten (18.4 mg/100 g dry wt) and the maximum content of all metabolites (64.0 mg/100 g dry wt) in R. graveolens ssp. divaricata callus obtained on the 35th culture day were relatively low. However, maximum contents of xanthotoxin (136.8 mg/100 g dry wt), bergapten (210.4 mg/100 g dry wt.) and isopimpinellin (96.7 mg/100 g dry wt), and total content of all metabolites in R. graveolens shoots (520.8 mg/100 g dry wt) obtained on the 42nd culture day are interesting from a practical point of view.

Accumulation of biologically active furanocoumarins in Ruta graveolens ssp. divaricata (Tenore) Gams in vitro culture.

The study was designed to investigate dynamics of accumulation of five linear furanocoumarins and umbelliferone in stationary liquid cultures of Ruta graveolens ssp. divaricata (Tenore) Gams during 6-week growth cycles. The contents of individual metabolites in biomass increased 1.8-3.5 times while their total content rose 2.3 times. Maximum contents of xanthotoxin, bergapten and isopimpinellin (112.3, 76.2 and 84.0mg/100g d.w., respectively) and maximum total content of all metabolites (283.4 mg/100 g d.w.), obtained on 35th culture day, are interesting from practical point of view.

Medicinal plant biotechnology: the Apiaceae family as the example of rapid development.

This article reviews the results of research representative of different trends in biotechnological studies of the Apiaceae species. These species are a well-known source of many important herbal products. A number of investigations focus on the biosynthesis of secondary metabolites in plant in vitro cultures. There are also numerous reports concerning the methods of plant micropropagation, especially using somatic embryogenesis. The papers dealing with the usage of the enzymatic potential of plant cells cultured in vitro, in biotransformation processes are less copious. Several studies are related to genetic engineering, exploring mainly the possibility of plant transformation with Agrobacterium rhizogenes to obtain hairy root cultures. The present review demonstrates the significance of biotechnological methods in the studies of medicinal plants taking species of the Apiaceae family as an example.

Coumarin compounds in Ammi majus L. callus cultures.

Callus cultures of Ammi majus L. (bishop's weed), Apiaceae, were maintained on variants of Linsmaier-Skoog's (L-S) medium differing in the content of the phytohormones, alpha-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP) (0.1-10.0 mg/l). The increments of callus tissue fresh weight showed considerable differences (1.4 bis 4.4-fold) during 4-week subcultures on the media tested. HPLC analysis revealed the presence of six compounds in the extracts of callus tissues, which are known metabolites in plants growing under natural conditions: the linear furanocoumarins psoralen, bergapten, xanthotoxin, isopimpinellin, imperatorin and their precursor umbelliferone. The total contents of coumarin compounds under examination showed marked differences dependent on the phytohormone concentrations in the medium. They ranged from 40.95 to 871.05 mg/100 g of dry weight. Imperatorin was the metabolite dominating among the furanocoumarins in calli from almost all tested variants of L-S medium (maximum content of 169.27 mg/100 g). Callus tissue cultured on one of the variants (0.1 mg/l NAA, 0.1 mg/l BAP) yielded in high content of xanthotoxin (145.33 mg/100 g). On the majority of media, umbelliferone was also accumulated at high quantities (maximum content of 536.29 mg/100 g). The medium containing 0.1 mg/l NAA and 0.1 mg/l BAP was the best for the accumulation of analysed coumarins. This medium favoured the formation of embryogenic callus. Xanthotoxin also dominated quantitatively among the furanocoumarins in the tested vegetative organs and fruits of the plant grown under natural conditions (leaves: 26.10 mg, roots: 5.55 mg, fruits: 3010.41 mg/100 g). Maximum contents of this metabolite in in vitro culture were many times higher than those found in vegetative plant organs but manifold lower than in fruits. On the other hand, maximum contents of imperatorin obtained in in vitro culture were many times higher in comparison with those detected in vegetative organs and fruits (leaves: 14.10 mg, roots: 3.30 mg, fruits: 94.70 mg/100 g of dry weight.). The A. majus L. callus culture, established in the course of the present experiments, can be considered a valuable model for studies of the biosynthesis of coumarin compounds, and a potential source of the psoralen 8-alkoxy derivatives imperatorin and xanthotoxin and their precursor umbelliferone.

Furanocoumarins in Pastinaca sativa L. in vitro culture.

Callus cultures of Pastinaca sativa L. (parsnip), Apiaceae, were cultivated on variants of Linsmaier-Skoog's medium, containing varying quantities (0.1-10.0 mg/l) of phytohormones: NAA-BAP and IBA-BAP which allowed to obtain 1.5-3-fold fresh biomass growth during 6-week subcultures. HPLC analyses showed that tissues cultured in vitro produced psoralen, bergapten, xanthotoxin, isopimpinellin and umbelliferone which are well known metabolites in plants growing under natural conditions. Total content of coumarins depended on the nature and quantity of phytohormones present in the medium, and ranged from 115.7 to 408.5 mg/100 g of the dry weight, isopimpinellin being the metabolite which dominated quantitatively (maximum content of 238.9 mg/100 g). Psoralen was also accumulated in callus tissues at considerable amounts (maximum content of 108.8 mg/100 g). This metabolite dominated in vegetative plant parts that have been analysed in our study (leaves, stems, roots) but its contents were lower than in the material from in vitro culture (48.9 mg/100 g 10.6 mg/100 g and 14.9 mg/100 g, respectively). Imperatorin was not detected in callus tissues although it dominated in the analysed fruits of the studied plant (200.0 mg/100 g). The best of the tested media in respect of promoting tissue biosynthetic capabilities was that which contained 3 mg/l NAA and 1 mg/l BAP. The studies showed that in vitro cultures of Pastinaca sativa L. can be a convenient model to study the biosynthesis of furanocoumarins and also a potential rich source of these compounds, particularly isopimpinellin.

In vitro cultures of higher plants and fungi as a potential source of bioactive metabolites.

The review presents the results of investigations conducted at the Chair of Pharmaceutical Botany, Collegium Medicum, Jagiellonian University, which demonstrated a prospects to obtain biologically active metabolites representative of many chemical groups (furanocoumarins, polysaccharides and lectins, indole compounds, carotenoids) in in vitro cultures of both higher plants and higher fungi (Macromycetes) (mycelial cultures). These cultures can be a potential, rich, new source of metabolites.

[Norms of EEG sleep patterns for healthy Polish population aged from 60 to 79 years]. / Normy wzorca EEG snu dla zdrowej populacji polskiej w wieku 60-79 lat.

Norms of sleep EEG patterns for a healthy population aged from 60 to 79 years are presented. The collected data were compared with data found in the literature.

Norms of EEG sleep patterns for a healthy Polish population aged from 20 to 59 years.

Norms of sleep EEG patterns for a healthy population aged from 20 to 59 years are presented from which it was concluded that this age group forms a relatively stable pattern during the ontogenesis of sleep. The collected data were compared with data found in the literature.

[Norms of EEG sleep patterns for healthy Polish population aged from 20 to 59]. / Normy wzorca eeg snu dla zdrowej populacji polskiej w wieku 20-59 lat.

Norms of sleep EEG patterns for a healthy population aged from 20 to 59 years are presented from which it was concluded that this age group forms a relatively stable pattern during the ontogenesis of sleep. The collected data were compared with data found in the literature.