Antistaphylococcal activity of Inula helenium L. root essential oil: eudesmane sesquiterpene lactones induce cell membrane damage.

The purpose of this study was to investigate the inhibitory/bactericidal activity and cell membrane effects of the hydrodistilled essential oil of Inula helenium L. roots against Staphylococcus aureus. Additionally, detailed chemical investigation was done in order to pinpoint the most active oil constituents and also the parts of these molecules responsible for their antimicrobial effect. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using the broth microdilution method. The membrane-active nature of this oil was investigated by measuring the culture turbidity, leakage of phosphates, and 260-nm-absorbing material, together with lysis of the exposed cells. Finally, the effect of the oil on the cells was visualized using scanning electron microscopy (SEM). The chemical composition of the essential oil was analyzed using gas chromatography-mass spectrometry (GC-MS) and preparative medium-pressure liquid chromatography (MPLC). Chemical modification of the oil was performed using catalytic hydrogenation (H(2), Pd/C) and reduction with NaBH(4). The MIC and MBC values were 0.01 µl mL(-1) and 0.02 µl mL(-1), respectively. Membrane damage was demonstrated through increased permeability (phosphates and nucleic acid leakage), followed by lysis of the exposed cells, captured on SEM images. The most active constituents were alantolactone, isoalantolactone, and diplophyllin. The essential oil showed very potent antistaphylococcal activity, with obvious membrane-damaging effects. Sesquiterpene lactones were found to be the most active principles of the oil, whose eudesmane core olefinic bonds, along with the α,ß-methylene-lactone ring, are essential structural parts responsible for the exhibited antimicrobial activity.

[New technology in maize breeding]. / Nove tehnologije u oplemenjivanju kukuruza.

Results obtained by several approaches in the application of Biotechnology in maize breeding are reviewed. RFLP technology in the determination of genetic variation; gene transfer by the use of different methods of gene delivery and the determination of gene integration. Three technologies for foreign gene introduction have been applied; injection of plasmid pRT100 neo into archesporial tissue before micro and macro sporogenesis, slightly modified pollen-tube pathway technology and dry seed incubation in plasmid DNA solution. NPTII gene integration was followed by dot-blot and Southern blot analysis of plant DNA of both T1 and T2 plants. Gene expression was analysed by neomycin phosphotransferase activity. Transformed plants contained the selective NPTII gene sequence in an active form. Bacterial gene integration induced several heritable changes of plant phenotype. As an important change, alteration of the flowering time has been used as a criterion for selection and plant propagation to keep transformed progeny. Besides plant genome transformation, endogenous bacteria living in different maize tissue were found. As a perspective approach for biotechnology application in maize breeding biological vaccine construction has been selected. Therefore, antagonistic effect of gram positive bacterial strains to several pathogenic fungi was investigated. Results obtained after in vivo experiments are discussed.