Tomas Vanek

Screening of some Siberian medicinal plants for antimicrobial activity

The antimicrobial activity of crude ethanolic extracts of 16 Siberian medicinal plants was tested against five species of microorganisms: Bacillus cereus, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. Of the 16 plants tested, 12 showed antimicrobial activity against one or more species of microorganisms. The most active antimicrobial plants were Bergenia crassifolia, Chelidonium majus, Rhaponticum carthamoides, Sanguisorba officinalis, and Tussilago farfara.

Biological remediation of explosives and related nitroaromatic compounds.

Nitroaromatics form an important group of recalcitrant xenobiotics. Only few aromatic compounds, bearing one nitro group as a substituent of the aromatic ring, are produced as secondary metabolites by microorganisms. The majority of nitroaromatic compounds in the biosphere are industrial chemicals such as explosives, dyes, polyurethane foams, herbicides, insecticides and solvents. These compounds are generally recalcitrant to biological treatment and remain in the biosphere, where they constitute a source of pollution due to both toxic and mutagenic effects on humans, fish, algae and microorganisms. However, relatively few microorganisms have been described as being able to use nitroaromatic compounds as nitrogen and/or carbon and energy source.The best-known nitroaromatic compound is the explosive TNT (2,4,6-trinitrotoluene). This article reviews the bioremediation strategies for TNT-contaminated soil and water. It comes to the following conclusion: The optimal remediation strategy for nitroaromatic compounds depends on many site-specific factors. Composting and the use of reactor systems lend themselves to treating soils contaminated with high levels of explosives (e.g. at former ammunition production facilities, where areas with a high contamination level are common). Compared to composting systems, bioreactors have the major advantage of a short treatment time, but the disadvantage of being more labour intensive and more expensive. Studies indicate that biological treatment systems, which are based on the activity of the fungusPhanerochaete chrysosporium or onPseudomonas sp. ST53, might be used as effective methods for the remediation of highly contaminated soil and water.Phytoremediation, although not widely used now, has the potential to become an important strategy for the remediation of soil and water contaminated with explosives. It is best suited where contaminant levels are low (e.g. at military sites where pollution is rather diffuse) and where larger contaminated surfaces or volumes have to be treated. In addition, phytoremediation can be used as a polishing method after other remediation treatments, such as composting or bioslurry, have taken place. Thisin-situ treatment method has the advantage of lower treatment costs, but has the disadvantage of a considerable longer treatment time. In order to improve the cost-efficiency, phytoremediation of nitroaromatics (and other organic xenobiotics) could be combined with bio-energy production. This requires, however, detailed knowledge on the fate of the contaminants in the plants as well as the development of efficient treatment methods for the contaminated biomass that minimise the spreading of the contaminants into the environment during post harvest treatment.

Nanoparticle-specific changes in Arabidopsis thaliana gene expression after exposure to ZnO, TiO2, and fullerene soot

The effect of exposure to 100 mg/L zinc oxide (nZnO), fullerene soot (FS) or titanium dioxide (nTiO(2)) nanoparticles on gene expression in Arabidopsis thaliana roots was studied using microarrays. After 7d, nZnO, FS, or nTiO(2) exposure resulted in 660 up- and 826 down-regulated genes, 232 up- and 189 down-regulated genes, and 80 up- and 74 down-regulated genes, respectively (expression difference>2-fold; p[t test]<0.05). The genes induced by nZnO and FS include mainly ontology groups annotated as stress responsive, including both abiotic (oxidative, salt, water deprivation) and biotic (wounding and defense to pathogens) stimuli. The down-regulated genes upon nZnO exposure were involved in cell organization and biogenesis, including translation, nucleosome assembly and microtubule based process. FS largely repressed the transcription of genes involved in electron transport and energy pathways. Only mild changes in gene expression were observed upon nTiO(2) exposure, which resulted in up- and down-regulation of genes involved mainly in responses to biotic and abiotic stimuli. The data clearly indicate that the mechanisms of phytotoxicity are highly nanoparticle dependent despite of a limited overlap in gene expression response.

In vitro anti-inflammatory activity of carvacrol: Inhibitory effect on COX-2 catalyzed prostaglandin E2 biosynthesisb

Possible anti-inflammatory effect of carvacrol was evaluated by in vitro cyclooxygenase-2 (COX-2) assay. Carvacrol inhibited production of prostaglandin E2 catalysed by COX-2 with an IC50 value of 0.8 μM what is practically the same concentration as the IC50 obtained for the standard inhibitors indomethacin and NS-398 with values of 0.7 μM and 0.8 μM, respectively. The COX-1 was inhibited approximately at the same rate (IC50 of 0.7 μM for carvacrol), which suggests non-selective inhibition of both enzyme isoforms. The results of the study demonstrate possible anti-inflammatory potential of this compound due to the inhibition of inducible COX-2 isoform.

Plant response to heavy metals and organic pollutants in cell culture and at whole plant level

In vitro cell culture experiments provide a convenient system to study basic biological processes, by which biochemical pathways, enzymatic activity and metabolites can be specifically studied. However, it is difficult to relate cell cultures, calli or even hydroponic experiments to the whole plant response to pollutant stress. In the field, plants are exposed to additional a-biotic and biotic factors, which complicate further plant response. Hence, we often see thatin vitro selected species perform poorly under soil and field conditions. Soil physical and chemical properties, plantmycorrhizal association and soil-microbial activity affect the process of contaminant degradation by plants and/or microorganisms, pointing to the importance of pot and field experiments.BackgroundIncreasing awareness in the last decade concerning environmental quality had prompted research into ‘green solutions’ for soil and water remediation, progressing from laboratoryin vitro experiments to pot and field trials.ObjectiveThis paper is a joint effort of a group of scientists in COST action 837. It represents experimental work and an overview on plant response to environmental stress fromin vitro tissue culture to whole plant experiments in soil.ResultsResults obtained fromin vitro plant tissue cultures and whole plant hydroponic experiments indicate the phytoremediation potential of different plant species and the biochemical mechanisms involved in plant tolerance. In pot experiments, several selected desert plant species, which accumulated heavy metal in hydroponic systems, succeeded in accumulating the heavy metal in soil conditions as well.Conclusions and RecommendationsIn vitro plant tissue cultures provide a useful experimental system for the study of the mechanisms involved in the detoxification of organic and heavy metal pollutants. However, whole plant experimental systems, as well as hydroponics followed by pot and field trials, are essential when determining plant potential to remediate polluted sites. Multidisciplinary research teams can therefore increase our knowledge and promote a practical application of phytoremediation.

Tolerance to, and Uptake and Degradation of 2,4,6-Trinitrotoluene (TNT) are Enhanced by the Expression of a Bacterial Nitroreductase Gene in Arabidopsis thaliana

Abstract Arabidopsis thaliana was transformed with a gene encoding a nitroreductase (NTR, E.C. 1.6.99.7) with activity against a wide range of nitroaromatic compounds. The gene was transferred from Escherichia coli by an Agrobacterium-mediated in planta method. The obtained seeds were sowed to produce T1 plants, and they were assayed for the integration of the transgene in the plant genome. Transgenic plants that were positive with the PCR analysis were self-pollinated to produce T2 generation plants. Seven lines obtained were assayed for the NTR activity. While the noil-transformed wild-type plants showed no detectable NTR activity, the enzyme activity of the transgenic plant lines was approx. 20 times higher. Using the line with the highest NTR activity, the phytoremediation characteristics of plants against 2,4,6-trinitrotoluene (TNT) was investigated. While the wild-type plants did not grow in the presence of 0.1 mᴍ TNT, the transgenic plants grew almost normally in this condition. The uptake of TNT by seedlings of transgenic plants increased by 7 to 8 times when they were floated on TNT solution. HPLC analysis showed that the peak due to TNT taken up into plant body was much smaller in the transgenic plants as compared with that of the wild type, and that a number of peaks attributable to the degradation products of TNT, including 4-amino-2,6-dinitrotoluene, were detected in the extract from the transgenic plants. This indicates that the expression of bacterial NTR improved the capability of plants to degrade TNT.

Evaluation of Antimicrobial and Anti-Inflammatory Activities of Seed Extracts from Six Nigella Species

Seed extracts from six species of the genus Nigella (Family Ranunculaceae)-Nigella arvensis, Nigella damascena, Nigella hispanica, Nigella nigellastrum, Nigella orientalis, and Nigella sativa-obtained by successive extraction with n-hexane, chloroform, and methanol, were tested for their antimicrobial activity against 10 strains of pathogenic bacteria and yeast using the microdilution method as well as for anti-inflammatory properties by in vitro cyclooxygenase (COX)-1 and COX-2 assay. Chemical characterization of active extracts was carried out including free and fixed fatty acid analysis. Comparison of antimicrobial activity showed that N. arvensis chloroform extract was the most potent among all species tested, inhibiting Gram-positive bacterial and yeast strains with minimum inhibitory concentration (MIC) values ranging from 0.25 to 1 mg/mL. With the exception of selective inhibitory action of n-hexane extract of N. orientalis on growth of Bacteroides fragilis (MIC = 0.5 mg/mL), we observed no antimicrobial activity for other Nigella species. Anti-inflammatory screening revealed that N. sativa, N. orientalis, N. hispanica, N. arvensis n-hexane, and N. hispanica chloroform extracts had strong inhibitory activity (more than 80%) on COX-1 and N. orientalis, N. arvensis, and N. hispanica n-hexane extracts were most effective against COX-2, when the concentration of extracts was 100 microg/mL in both COX assays. In conclusion, N. arvensis, N. orientalis, and N. hispanica seeds, for the first time examined for antimicrobial and anti-inflammatory effects, revealed their significant activity in one or both assays.

Accumulation and Transformation of Sulfonated Aromatic Compounds by Rhubarb Cells (Rheum palmatum)

ABSTRACT Sulfonated aromatic compounds are released into the environment in large amounts: the main sources of these recalcitrant pollutants are anionic detergents, dyestuffs, and their byproducts, such as sulfonated anthraquinones. Different derivatives of anthraquinones occur naturally in several plant species such as rhubarb. Therefore, this plant could possess enzymes able to accept sulfonated anthraquinones as substrates. Previously, it has been shown that cultured cells isolated from Rheum palmatum are able to efficiently accumulate and transform different mono- and disulfonated anthraquinones. Because rhubarb is a hardy plant species, it may prove a promising candidate in developing new biological processes to decontaminate effluents containing recalcitrant xenobiotics. The ability of rhubarb cells to accumulate and transform other sulfonoaromatics was investigated in this study. Results obtained show that cultured rhubarb cells were able to efficiently accumulate 2-chloro-5-nitro-benzene sulfonate...

Production of saponins from Panax ginseng suspension and adventitious root cultures

Biomass growth and ginsenoside production in cell suspension and adventitious roots of Panax ginseng C.A. Meyer cultures cultivated both in Erlenmayer flasks and a 3 dm3 bioreactor were studied. The maximum content of ginsenosides was found in the suspension culture cultivated in the bioreactor (4.34 % dry mass), however the saponin content was limited to two major ginsenosides, Rb1 and Rg1. The production of ginsenosides in adventitious roots was lower (1.45 or 1.72 % dry mass), nevertheless, the full range of ginsenosides was detected.

Biotransformation of (S)-(−)- and (R)-(+)-limonene using Solanum aviculare and Dioscorea deltoidea plant cells

Abstract (S)-(−)- and (R)-(+)-limonene was transformed to carvone via corresponding cis- and trans-carveol using Solanum aviculare and Dioscorea deltoidea plant cells. Both carveols and carvone formed were racemic in all biotransformations.