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Two xanthones from Polygala paniculata and confirmation of the 1-hydroxy-2,3,5-trimethoxy-xanthone at trace level by HRGC-MS.

Polygala paniculata L. yielded the xanthones 1-hydroxy-5-methoxy-2,3-methylenedioxyxanthone (1) and 1,5-dihydroxy-2,3-dimethoxyxanthone (2), together with coumarin murragatin and flavonol rutin. Their structures were established by chemical and spectroscopic methods (EIMS, IR, 1H and 13C NMR, NOE difference spectroscopy). By posterior analysis of an apolar crude extract using high resolution gas chromatography coupled to mass spectrometry (HRGC-MS) it was possible to characterize two sterol (spinasterol and delta25-spinasterol) and the minor 1-hydroxy-2,3,5-trimethoxyxanthone (3). Thus, the xanthone 3 was confirmed through of co-injection HRGC-MS of the respective extract with a certified standard obtained by methylation of 2 with diazomethane.

Chromatography, Gas↗

Reconstructing the evolutionary history of paralogous APETALA1/FRUITFULL-like genes in grasses (Poaceae).

Gene duplication is an important mechanism for the generation of evolutionary novelty. Paralogous genes that are not silenced may evolve new functions (neofunctionalization) that will alter the developmental outcome of preexisting genetic pathways, partition ancestral functions (subfunctionalization) into divergent developmental modules, or function redundantly. Functional divergence can occur by changes in the spatio-temporal patterns of gene expression and/or by changes in the activities of their protein products. We reconstructed the evolutionary history of two paralogous monocot MADS-box transcription factors, FUL1 and FUL2, and determined the evolution of sequence and gene expression in grass AP1/FUL-like genes. Monocot AP1/FUL-like genes duplicated at the base of Poaceae and codon substitutions occurred under relaxed selection mostly along the branch leading to FUL2. Following the duplication, FUL1 was apparently lost from early diverging taxa, a pattern consistent with major changes in grass floral morphology. Overlapping gene expression patterns in leaves and spikelets indicate that FUL1 and FUL2 probably share some redundant functions, but that FUL2 may have become temporally restricted under partial subfunctionalization to particular stages of floret development. These data have allowed us to reconstruct the history of AP1/FUL-like genes in Poaceae and to hypothesize a role for this gene duplication in the evolution of the grass spikelet.

Amino Acid Sequence↗

Research into microscopic structure and essential oils of endemic medicinal plant species Satureja subspicata Bartl. ex Vis. (Lamiaceae).

In this study we looked into the cells and histological organization of leaves (Saturejae folium) as well as a phyto-chemical composition of overground parts (Saturejae herba) of endemic species Satureja subspicata Bartl. ex Vis. (Lamiaceae) collected during year 2003 on south slopes of mountain Velez in Herzegovina. Microscopic organization was analyzed in wet slides using light microscope. Estimation of stomata index was done according to Ph. Yug. IV. Chemical composition of overground material extracts was determined by thin layer chromatography (TLC) using thymol as a reference. In our research we found the following: Leaf structure of the analyzed species Satureja subspicata points at numerous specificities in anatomical and histological sense. In histological sense, leaf is of ventral type, with differentiated upper and lower epidermis and palisade and spongy tissue in between. Stoma index assigned according to Ph. Yug. IV leads to a conclusion that it is the case of diastitic stomata, which is common feature of most species from Lamiaceae family. Comparative qualitative analysis of essential oils in species Satureja subspicata showed similarities with other species from Lamiaceae family such as Thymus L. (thymol). In fact, we found more common substances that are part of the species Satureja montana L. extract, but in different concentrations.

Bosnia and Herzegovina↗

A trait-based test for habitat filtering: convex hull volume.

Community assembly theory suggests that two processes affect the distribution of trait values within communities: competition and habitat filtering. Within a local community, competition leads to ecological differentiation of coexisting species, while habitat filtering reduces the spread of trait values, reflecting shared ecological tolerances. Many statistical tests for the effects of competition exist in the literature, but measures of habitat filtering are less well-developed. Here, we present convex hull volume, a construct from computational geometry, which provides an n-dimensional measure of the volume of trait space occupied by species in a community. Combined with ecological null models, this measure offers a useful test for habitat filtering. We use convex hull volume and a null model to analyze California woody-plant trait and community data. Our results show that observed plant communities occupy less trait space than expected from random assembly, a result consistent with habitat filtering.

Ecosystem↗

Application of the in combo screening approach for the discovery of non-alkaloid acetylcholinesterase inhibitors from Cichorium intybus.

Because of the direct correlation of cholinergic deficit and the severity of dementia, Alzheimer's disease is preferentially treated with acetylcholinesterase (AChE) inhibitors to supplement the acetylcholine level. In this study we focused on non-alkaloid AChE inhibitors from natural sources in order to discover new lead structures. In the course of in vitro extract screening of Tyrolean plants using an enzyme assay with Ellman's reagent, the dichloromethane extract of chicory roots (Cichorium intybus L.) showed a pronounced inhibitory effect on AChE. At a concentration of 1 mg extract/ml an inhibition of 70% was measured. Based on a 3D multi-conformational molecular-structure database consisting of secondary metabolites from C. intybus known from the relevant literature, virtual screening filtering experiments were conducted using both a feature-based pharmacophore model and a docking procedure. Some low molecular weight sesquiterpenoids exhibited distinct interactions with the pharmacophore model. In order to verify the applicability of this computer-aided strategy, an activity-guided fractionation of the chicory root extract was performed, which resulted in the isolation of two sesquiterpene lactones, 8-deoxylactucin and lactucopicrin, showing significant and dose-dependent inhibitory activity on AChE (IC(50) of 308.1 microM [CI(95) 243.9 - 405.3 microM] and 150.3 microM [CI(95) 100.8 - 188.1 microM], respectively). The two isolates were correctly predicted within the virtual screening process which corroborates the potential of the computer-assisted in combo screening approach for the discovery of the anti-cholinesterase compounds from C. intybus.

Acetylcholinesterase↗

Evaluation of primary and secondary treated and disinfected wastewater irrigation of tomato and cucumber plants under greenhouse conditions, regarding growth and safety considerations.

Tomato and cucumber seedlings were distributed into 10 groups (five for each plant) of 15 plants each. The plants were irrigated for 10 weeks with primary treated wastewater (group A), secondary treated wastewater (group B), chlorinated secondary treated wastewater (group C), a fertilizer dilution (group F), and tap water (group M). All precautions were taken to secure that there was no direct contact between the wastewater and the edible portions of the crops. During this period and on a weekly basis, the height and number of leaves was monitored, while, at the end, the dry weight of leaves, stems, and roots for each plant of each group was measured. Based on these growth parameters, both types of plant in groups A and F recorded the most significant development compared to the other three groups. The plants irrigated with tap water recorded the smallest development, in every case. Plants in groups B and C were similar, with a slight (but not significant) superiority for the plants irrigated with secondary treated wastewater, probably as a result of some phytotoxic effects of residual chloride in the chlorinated wastewater. The presence of nutrients and specifically nitrogen in the various solutions explains the differences satisfactorily. The vegetables grown on the plants of each group were harvested, and their surface tissue analyzed for total coliforms (TC) and enterococci (EC). Tomatoes grown on the plants of groups A and B recorded the highest values for TC, with 505 and 490 cfu/g, respectively, whereas, for cucumbers, those values were 342 and 450 cfu/g, respectively. Enterococci were detected on the surface of harvested vegetables from groups A and B, but the small number of cases and their random character cannot support any strong relations between the used wastewater and their presence. The TC values in group C were very low, far lower than those if group F. No EC were found in either group C or group F. These primary results suggested that irrigation with appropriate disinfected wastewater, even of such high-risk cultivations of vegetables eaten raw, should not be discarded completely as unsafe, but be reconsidered and studied further. However, the use of undisinfected wastewater in such greenhouse cultivations, where all safety precautions have been taken to prevent any contact of the fruits with the soil or the wastewater, does not prove to be 100% safe.

Consumer Product Safety↗

Cytotoxicity of water extracts from leaves and branches of Philadelphus coronarius L.

Philadelphus coronarius L. is big, leggy and deciduous old-fashioned shrub known for its fragrant white flowers in the late spring. Some members of genus Philadelphus L. are known for their antibacterial, antiradical and immunomodulatory effects. Therefore, these herbs represent prospective sources for the isolation of active substances with desired effects. We have investigated the cytotoxicity effects of water extracts from leaves and branches of Philadelphus coronarius L. (Hydrangeaceae). A431 cells (human skin carcinoma cell line) and the human breast adenocarcinoma cell line (MCF-7) were treated with various doses of individual extracts (0,1-100 microg dry matter/ml) for 24 h and 72 h. The highest toxic effects of both plant parts extracts were observed on MCF-7 cells regardless the time of treatment. Cells A431 were less sensitive to toxic effects of leaves and branches extracts but the time dependence was present with the tendency of increased toxicity after chronic treatment. There were no differences in the extent of toxic effects between branches and leaves extracts. The results obtained so far will provide the basis for the future studies with isolated active substances from these extracts.

Antineoplastic Agents, Phytogenic↗

[Pharmacognostic identification of Ampelopsis grossedentata].

Pharmacognostic identification of Ampelopsis grossedentata was carried out by studying its microscopic characteristics, UV spectra and TLC. The results may provide a scientific basis for the comprehensive development and utilization of the drug.

Ampelopsis↗

[Studies on calluses induced from various explants of Cistanche deserticola].

Calluses were induced from various explants (flower, stem, bud and squama) of Cistanche deserticola Y.C.Ma and compared with each other. The result showed that the inducement rate of ovary and stem were higher than that of other explants which the inducement time was short and the characters of calluses were good. The effect of basal culture medium added hormone on callus reproduction was also studied. The result showed that B5 culture medium added 1 mg/L IBA and 2 mg/L 6-BA was optimal.

Cistanche↗

[Study on extraction of active ingredients from Andrographis paniculatal using the orthogonal experiment with supercritical carbon dioxide].

This paper studied the extraction technology of active ingredients from Andrographis paniculatal using the orthogonal experiment with supercritical CO2. The active ingredients of this extract had higher purity and more stable quality than those came from conventional extract technology. And the target ingredients, dehydrated andrographolidume and andrographolidume, had higher content. Furthermore this method had shorter technology process and saved a great deal of solvent than conventional ethanol extraction, which was exactly suitable for industrialization manufacture. The optimization condition was: extractor pressure 25 MPa, extractor temperature 46 degrees C, separator I pressure 6 MPa, separator I temperature 65 degrees C, separator II pressure 6 MPa, separator II temperature 45 degrees C, CO2 rate of flow 40 kg/h.

Andrographis↗

[Role of trophic and hormonal factors in exogenous regulation of the formation of reproductive organs in yellow lupine (Lupinus luteus L.)].

During the formation and ripening of beans in yellow lupine, their intactness may be increased by treatment of the plants with the microelement molybdenum or the growth factor 24-epibrassinolide. Early stages of the effects of either factor involved activation of various metabolic pathways; eventually, the supply of fruit elements with free amino acids was improved and the seed productivity of the plants, increased.

Brassinosteroids↗

[Determination of cichoric acid in Echinacea purpuea].

OBJECTIVE: To analyze the natural change rule of active components of E. purpuea by measuring content of cichoric acid. METHOD: Reverse HPLC method was used. RESULT: The maximum cichoric acid content of the roots occured in seedling age of May, and that of the flowers occured in blooming stage of mid July, but cichoric acid in stems was generally low anyway. The maximum content of cichoric acid in the plant above ground occured in the blooming stage of mid July. CONCLUSION: The measuring method of content of cichoric acid is successful and reliable. The optimum stage of harvest in Echinacea purpuea should be guided by natural change rule of cichoric acid content.

Caffeic Acids↗

[Studies on the chemical constituents from Myristica yunnanensis Y. H. Li].

OBJECTIVE: To study the chemical constituents of Myristica yunnanensis. METHOD: Compounds were isolated using silica gel and reversed-phase C18 column chromatography, their structure were identified by means of spectral method. RESULT: Five known compounds were identified as otobain(I), beta-sitosterol(II), ergosterol(III), afzelin(IV) and quercetrin(V). CONCLUSION: Compounds IV and V were isolated first from Myristica.

Flavonoids↗

[Chemical constituents from Salix oritrepha Schneid].

OBJECTIVE: To study chemical constituents of the above ground part of Salix oritrepha. METHOD: Using chromatography to isolate compounds and their structure were identified by physical, chemical and spectral techniques. RESULT: Four compounds were isolated and elucidated as beta-sitosterol, beta-sitosteryl-3-O-glucoside, friedelin, luteolin-7-O-glucoside. CONCLUSION: All of them were isolated from S. oritrepha for the first time. Friedelin from the plants of genus Salix for the first time.

Flavonoids↗

[Changes of dehydroandrographolide's contents of andrographis tablet in the process of production].

OBJECTIVE: To recognize changes in the contents of ingredients of Andrographis Tablet in the process of production. METHOD: Adopting TLCS, TLC, HPLC to detect effective contents of ingredients which are produced in every stage of process of Andrographis Table's production. RESULT: Handling with the fresh Herba Andrographis according to current pharmacopeoia's technology, it showed that only dehyandrographolide can be detected. It indicated that the main factor that leads to chemical change is the heating process in the process of production. CONCLUSION: Avoiding heating treatment or reducing heating treatment time is the main factor to protect the effective ingredients.

Andrographis↗

[Studies on chemical constituents of Sorbaria sorbifolia].

OBJECTIVE: To study the chemical constituents in the medicative part of Sorbaria sorbifolia. METHOD: By means of silica gel re-chromatography and thin layer chromatography, chemical constituents from the CHCl3 and EtOAc extract of the medicative part of S. sorbifolia were studied and their structures were identified by their physical-chemical constants, spectral analysis and comparison with the authorized samples. RESULTS: Eight compounds were obtained and identified as follows: noreugenin (I), wogonin (II), 5,7,3',4'-tetrahydroxy-3-methoxyflavone (III), protocatechuic acid (IV), benzoic acid (V), p-hydroxy benzoic acid (VI), emodin (VII) and daucosterol (VIII). CONCLUSION: All of the compounds were isolated from this genus for the first time.

Flavanones↗