Sequence analysis of 16S rRNA, gyrB and catA genes and DNA-DNA hybridization reveal that Rhodococcus jialingiae is a later synonym of Rhodococcus qingshengii.

The results of 16S rRNA, gyrB and catA gene sequence comparisons and reasserted DNA-DNA hybridization unambiguously proved that Rhodococcus jialingiae Wang et al. 2010 and Rhodococcus qingshengii Xu et al. 2007 represent a single species. On the basis of priority R. jialingiae must be considered a later synonym of R. qingshengii.

Investigation of mineral water springs of Miercurea Ciuc (Csíkszereda) region (Romania) with cultivation-dependent microbiological methods.

Water samples of ten mineral water springs at Miercurea Ciuc (Csíkszereda) region (Romania) were examined during 2005-2006 using cultivation-dependent microbiological methods. The results of standard hygienic bacteriological tests showed that the Hargita Spring had perfect and five other springs had microbiologically acceptable water quality (Zsögöd-, Nagy-borvíz-, Taploca-, Szentegyháza- and Lobogó springs). The water of Borsáros Spring was exceptionable (high germ count, presence of Enterococcus spp.).Both standard bacteriological and molecular microbiological methods indicated that the microbiological water quality of the Szeltersz-, Nádasszék- and Délo springs was not acceptable. Bad water quality resulted from inadequate spring catchment and hygiene (low yield, lack of runoff, negligent usage of the springs, horse manure around the spring).The 16S rRNA gene-based identification of strains isolated on standard meat-peptone medium resulted in the detection of typical aquatic organisms such as Shewanella baltica, Aeromonas spp., Pseudomonas veronii, Psychrobacter sp,. Acinetobacter spp. and allochthonous microbes, like Nocardia, Streptomyces, Bacillus, Microbacterium , and Arthrobacter strains indicating the impact of soil. Other allochthonous microbes, such as Staphylococcus spp., Micrococcus sp., Lactococcus sp., Clostridium butyricum, Yersinia spp., Aerococcus sp., may have originated from animal/human sources.

Cytotoxic activities of Stachys species.

The cytotoxic activity of Stachys plants and of aucubin and harpagide against MCF7-breast adenocarcinoma, HeLa-cervix adenocarcinoma, A431-skin carcinoma of epithelial origin is reported in this study. Cisplatin and doxorubicin were use as reference compound.

Comparison of antioxidant activity in enzyme-independent system of six Stachys species.

The antioxidant activity of six Stachys species native to Hungary in an enzyme-independent lipid-peroxidation system was examined. The methanolic extracts were found to be more effective than the controls alpha-tocopherol succinate and ascorbic acid. The following components were determined by spectrophotometric analysis: hydroxycinnamic acid derivatives, phenols content and flavonoids.

5-Alpha- and 5-beta-2-deoxyintegristerone A, a 5-alpha and 5-beta isomer pair of ecdysteroids isolated from the Silene genus.

5-Alpha-2-deoxyintegristerone A and 5-beta-2-deoxyintegristerone A were isolated from the aerial parts of Silene italica ssp. nemoralis (Waldst. and Kit.) Nyman using a specific combination of absorption column chromatography, preparative thin-layer chromatography and preparative HPLC. Both normal-phase and reversed-phase modes of HPLC were employed for isolation. Structural elucidation of 5-alpha-2-deoxyintegristerone A was completed by X-ray diffraction. Both 5-alpha-2-deoxyintegristerone A and 5-beta-2-deoxyintegristerone A were firstly isolated from this plant. We propose that 5-alpha-2-deoxyintegristerone A is not an artifact but an integral part of the ecdysteroid spectrum of Silene italica ssp. nemoralis (Waldst. and Kit.) Nyman.

Antioxidant activity of leaves of Salvia species in enzyme-dependent and enzyme-independent systems of lipid peroxidation and their phenolic constituents.

The protective effects of eleven Salvia species native to Europe against enzyme-dependent and enzyme-independent lipid peroxidation were evaluated. The 50% aqueous methanolic extracts of the leaves of all tested plants were found to be more effective than the positive control alpha-tocopherol acid succinate. The extracts of S. candelabrum, S. ringens, S. tomentosa, S. nemorosa, and S. glutinosa displayed considerable concentration-dependent antioxidative effects that were comparable to those of the medicinal and aromatic plant S. officinalis. The concentrations of flavonoids, hydroxycinnamic acids and total phenolic compounds in each extract were quantified with the aim of clarifying the connection between activity and chemical composition.

[Structural diversity of ecdysteroids of Lychnis flos-cuculi]. / A Lychnis flos-cuculi ekdiszteroidjainak szerkezeti diverzitása.

Eleven ecdysteroids have been isolated from Lychnis floscuculi; we are the first who report eight ecdysteroids of the eleven compounds in this plant. Two of these ecdysteroids, dihydrorubrosterone and 20-hydroxyecdysone 3-acetate are newly discovered natural products. The success of isolation of these new ecdysteroids has been based on the use of separation methods in a proper order; these separation procedures were completing each others. At the beginning steps of isolation simple separation methods were used, such a solvent-solvent distribution and fractionated precipitation. Two third of the contaminants were removed thereby. High capacity low resolution methods were used then, such as classical adsorption column chromatography and preparative thin-layer chromatography. The major component (20-hydroxyecdyssone) and certain minor ecdysteroids (polypodine B and rubrosterone) were isolated in pure form here. Purification of the further minor components (poststerone, 2-deoxy-20-hydroxyecdysone, vitikosterone E, dihydrorubrosterone, makisterone A, taxisterone, 20-hydroxyecdysone 2-acetate, 20-hydroxyecdysone 3-acetate) required HPLC and other absorption chromatographic methods. Our recent separation scheme means a generally applicable guiding principle for isolation of any plant ecdysteroid, major and minor alike. Structural identification of the known ecdysteroids was based on their spectral data and that of their literature information. Structural elucidation of 20-hydroxyecdysone 3-acetate was done by the help of a standard component prepared by acetylation of 20-hydroxyecdysone. From the mixture of seven acetates the corresponding compound (20-hydroxyecdysone 3-acetate) was isolated, and used for identification. Structural diversity of ecdysteroids of Lychnis flos-cuculi is evaluated, and a tentative explanation is introduced for the formation and biosynthesis of the versatility of phytoecdysteroids.

Isolation of 5 alpha- and 5 beta-dihydrorubrosterone from Silene otites L. (Wib).

5 alpha-Dihydrorubrosterone (2 beta, 3 beta, 14 alpha, 17 beta-tetrahydroxy-5 alpha-androst-7-ene-6-one), a new 19-carbon 5 alpha-ecdysteroid, was isolated together with its 5 beta counterpart from the aerial parts of Silene otites L. (Wib.) (Caryophyllaceae) by a combination of solvent partition, low-pressure column chromatography, thin-layer chromatography (normal-phase and reversed-phase) and finally HPLC. Mass spectrometry and nuclear magnetic resonance spectroscopic procedures were used for compound characterization.

Protective effects of the aerial parts of Salvia officinalis, Melissa Officinalis and Lavandula angustifolia and their constituents against enzyme-dependent and enzyme-independent lipid peroxidation.

The antioxidant effects of aqueous methanolic extracts from three medicinal Lamiaceae species were investigated in enzyme-dependent and enzyme-independent lipid peroxidation systems. All these extracts caused a considerable concentration-dependent inhibition of lipid peroxidation. Phenolic components present in the plant extracts were evaluated for antioxidant activity and were found effective in both tests. Their concentrations in each extract were determined by TLC-densitometry.

Jatrophane diterpenoids from Euphorbia peplus.

From the pro-inflammatory active extract of Euphorbia peplus, a new diterpene polyester (1) based on the jatrophane skeleton was isolated together with the known compounds 2-5. The irritant activities of some jatrophane diterpenes (2, 3 and 6-9) were also investigated: only compound 2 was found to exert a weak pro-inflammatory activity on mouse ear.

[Components of Serratula species; screening for ecdysteroid and inorganic constituents of some Serratula plants]. / Serratula fajok 20-hidroxiekdizon és szervetlen elem tartalmának vizsgálata.

Ecdysteroid and inorganic components were analyzed from several plant species belonging to Caryophyllaceae family, such as in the cases of Serratula tinctoria, Serratula wolffii, Serratula coronata (20-hydroxyecdysone and inorganic components) and Jurinea mollis, Serratula gmellinii and Leuzea carthamoides (inorganic components, only). The 20-hydroxyecdysone content was determined using thin-layer chromatography after a simple clean up that had been performed by solid-phase extraction. Inorganic constitutents were determined using either ICP or flame photometry. Vegetation dependence of both 20-hydroxyecdysone and inorganic elements was studied. Serratula coronata shows remarkable high 20-hydroxyecdysone (2.3%, in April) and the studied Serratula plants gave a minimum of 20-hydroxyecdysone in June. Favorable period for harvesting is suggested as July and August, through the blossoming of these plants. Potassium and 20-hydroxyecdysone content gave a similar tendency considering their vegetation dependence, while the magnesium content moved toward opposite direction. The calcium contents of Serratula tinctoria, Serratula wolffii and Serratula coronata were found between 2.2% and 3.8%, which values are high relative to the other medicinal plants. At the same time, the Cu content of the ecdysteroid producing (and screened) Caryophyllaceae plants is low. The Fe, Mn and Mg contents of Serratula coronata are high, even higher than that of the Leuzea carthamoides. Our results have suggested the importance of analysis and control of inorganic constituents of crude plant extracts used for medicinal and recreational purposes.

A novel lathyrane diterpenoid from the roots of euphorbia lathyris

A new lathyrane diterpene (1) has been isolated and characterized from a CH2Cl2 extract of the roots of Euphorbia lathyris. Detailed spectral analysis revealed that the structure of 1, including relative stereochemistry, is that of a diester of a hitherto unknown, polyfunctional diterpene parent alcohol.

[Densitometric method for parallel measurement of rosmarinic and caffeic acids in plant samples]. / Denzitometriás módszer növényminták rozmaring- és kávésav tartalmának együttes mérésére.

In the last few years, the role of free radicals in the promotion of cell aging and tumour growth has focused attention on phenolic compounds. Rosmarinic acid (RA) and caffeic acid (CA), as major components of tannins, commonly occur together in many species of the Lamiaceae. A TLC-densitometric method was introduced for the parallel determination of RA and CA. Experiments were performed to establish the optimal measurement parameters related to the excitation wavelength, colour stability, linearity of the calibration curves, sensitivity and reproducibility. In the wavelength range 290-330 nm, it was found that the emission peak areas exhibit a maximum at 325 nm. Therefore, the densitometric evaluations of these tannins were made at 325 nm. The two compounds behaved very similarly as regards colour stability. The changes during the 3 hours of observation were not strong, as reflected by coefficients of variation of 8.20% for RA and 5.40% for CA. The calibration plots demonstrating the relation between the spotted amount and the measured peak area were found to include 2 intersecting linear sections, in the intervals 0-0.40 microgram and 0.40-1.20 micrograms for RA and 0-0.08 microgram and 0.08-0.30 microgram for CA. The correlation coefficients ranging between 0.991 and 0.998. The lower limits of detection were 1 ng for RA and 0.1 ng for CA. The reproducibility proved to be very good: the 10 times repeated densitometric estimation of the same spot yielded a coefficient of variation of 0.34% for RA and 0.36 for CA. The application of TLC permits utilization of the fluorescence of both phenols, often without the need for prior purification. This fluorescence measurement makes the detection extremely sensitive. The densitometric quantification allows the routine analysis of many samples in a short time with reasonable accuracy. The practical application of the elaborated method will be demonstrated on the leaves of 5 Salvia species. The results are shown in table I.

[Macrocyclic diterpene polyesters of the jatrophane type from Euphorbia esula]. / Jatrofánvázas makrociklusos diterpén-poliészterek az Euphorbia esula-ból.

Three new jatrophane diterpenes, esulatin A, B and C (1-3) were isolated and characterized from the whole, undried plant of Euphorbia esula. By means of spectral analysis, the structures were established as penta- and heptaesters of hitherto unknown, polyfunctional diterpene parent alcohols. Esulatin A (1) and C (3) are the diterpenoids with the highest degree of esterification identified to date from the family Euphorbiaceae.

[Investigation of variation of the production of biological and chemical compounds of Hyssopus officinalis L]. / Hyssopus officinalis L. produkcióbiológiai és kémiai változékonyságának tanulmányozása.

Hyssopus officinalis L. (Lamiaceae family) has been cultivated in Central Europe for a long time. This essential oil containing species serves not only as spice but in many countries including Hungary, it is used as a folk medicine against certain respiratory diseases. Despite this fact, little is known about the variation of its productivity under Central European climatic conditions. The cultivated populations of hyssop can be characterised by a significant heterogenity. In the course of its breeding the uniformity of flower colour (e.g. blue form), and increase in the oil content are the main achievable purposes. The purpose of this work was to investigate both the variability of strains of different crigin and the time-dependent variations of its production parameters. The optimum of phytomass was obtained at the beginning of July. The essential oil content as well as compounds of the non volatile fractions were also investigated. The non volatile fractions for rosmarinic, caffeic acids were analysed mainly by TLC and densitometry. Both compounds were present in all samples and they are suitable for the characterisation of the plant. The essential oils were gained with Water Steam Distillation (WSD) and Supercritical Fluid Extraction (SFE) with CO2. The oils were analysed by GC, GC-MS techniques. In the essential oil composition of the populations studied significant heterogenity could be observed. In the case of applying SFE extraction the oil composition is more uniform, similarly to the obtained by WSD adding hexane. The heterogenity can be experienced in the offsprings, too. If only the main four components (beta-pinene, limonene, pinocamphone, isopinocamphone) are regarded, among the offsprings clear and mixed lines alike can be found. Results of these experiments justify the necessity and usefulness of selection which is going on.

Isolation and Structural Elucidation of Two Plant Ecdysteroids, Gerardiasterone and 22-Epi-20-hydroxyecdysone

Two minor plant ecdysteroids, 22-epi-20-hydroxyecdysone (1) and gerardiasterone (2), were isolated from Serratula tinctoria L. (Compositae). The first compound, a new natural product, was characterized by an unusual stereochemistry at C-22 (i.e., 22S). The second compound was identified as (20R,23S)-20,23-dihydroxyecdysone, a compound previously isolated from the Zooanthid Gerardia savaglia.

New minor ecdysteroids from Silene otites (L.) Wib.

Six minor new ecdysteroid components have been isolated from Silene otites (L.) Wib. by a combination of chromatographic methods. Three of them (2-deoxy-20-hydroxyecdysone 3,22-diacetate, 5 alpha-2-deoxy-20-hydroxyecdysone 3-acetate, and 2-deoxy-20-hydroxyecdysone 3-crotonate) are new natural products.

[Chemical tests with Marrubium species. Official data on Marubii herba in Pharmacopoeia Hungarica VII]. / Marrubium fajok növénykémiai vizsgálata Adatok a VII. Magyar Gyógyszerkönyv Marrubii herba cikkelyéhez.

About 40 species of the Marrubium genus (Lamiaceae) are known of which 2 species (M. vulgare L. and M. peregrinum L.) and one hybrid (M. x paniculatum Desr.) can be found as native plants in Hungary. The above-ground parts of M. vulgare L. are official in Hungarian Pharmacopoeia VII. Active substances in Marrubii herba are labdane-structured bitter materials. Although the presence of furanic labdane diterpenes in the plant is known, the pharmacopoeia gives only microscopic tests, qualitative tests (for other parts of the plant and foreign organic matter) for the bitter value of Marrubii herba. We have examined the main terpenoid substances isolated with column, gel and preparative layer chromatography. Structure elucidations were performed by means of UV, mass and NMR spectroscopy. We have compared the changes in terpenoid-type compounds (premarrubiin and marrubiin) in plants during the vegetation period; in different Marrubium species and in the different extractions of horehound by means of thin layer chromatography and densitometry. By reason of our results we propose qualitative and quantitative chemical tests for the paragraph of Marrubii herba in Pharmacopoeia Hungarica VII.