Band 61, Heft 3-4

The antimicrobial activity of three Salvia species, i.e. S. santolinifolia, S. hydrangea and S. mirzayanii, essential oils were investigated. The essential oils were obtained from the aerial parts of plants and analyzed by GC-MS. The main constituents of aforementioned species were α-pinene (72.4%), β-pinene (6.6%) and limonene (5.3%); β-caryophyllene (25.1%), 1,8-cineol (15.2%) and caryophyllene oxide (11.5%); α-terpinenyl acetate (22.6%), 1,8-cineol (21.2%) and linalool (8.9%), respectively. Bioassays exhibited that the property of the oil of S. myrzayanii was superior to others. The antimicrobial activity of essential oil from Salvia species may well be due to the presence of synergy between six tested compounds (linalool, 1,8-cineol, α-pinene, β-pinene, β-caryophyllene and limonene) and other constituents of the oils with various degrees of antimicrobial activity. Among these, linalool and 1,8-cineol had the highest antimicrobial activity.

Separation and isolation of the two main compounds suaveolol and methyl suaveolate from leaves of chichinguaste (Hyptis suaveolens Poit., Lamiaceae) could be achieved by means of repeated column chromatography and repeated preparative thin layer chromatography. Their chemical structures were approved by MS, 1 H NMR, 13 C NMR and 2D-NMR experiments. The anti-inflammatory activity of the two compounds was tested for the first time as inhibition of croton oil-induced dermatitis of the mouse ear. Suaveolol and methyl suaveolate showed nearly the same dose-dependent topical anti-inflammatory activity, only two to three times lower than that of the reference drug indomethacin. The anti-inflammatory properties of these compounds could contribute to the antiphlogistic activity of extracts of Hyptis species and confirm the rational use of Hyptis suaveolens extracts in dermatological diseases.

Bioassay-directed fractionation of the cytotoxicity active fraction of the whole plant from Solanum lyratum led to the isolation of a new steroidal saponin, diosgenin 3-O-β-ᴅ-glucopyranosiduronic acid methyl ester (2), as well as four known compounds, diosgenin (1), diosgenin 3-O-β-ᴅ-glucopyranosiduronic acid (3), diosgenin 3-O-α-ʟ-rhamnopyranosyl-(1→2)-β-ᴅ-glucopyranosiduronic acid (4), diosgenin 3-O-α-ʟ-rhamnopyranosyl-(1→2)-β-ᴅ-glucuroniduronic acid methyl ester (5). The structures of the isolated compounds were elucidated on the basis of their spectral data and chemical evidences. Compound 1 was isolated for the first time from this plant, and compound 3 was isolated as a new natural product. Cytotoxic activities of the isolated compounds were evaluated and the cytotoxicities of compounds 2-5 reported for the first time.

From roots of Hyptis martiusii Benth. two tanshinone diterpenes were isolated, the new 7β-hydroxy-11,14-dioxoabieta-8,12-diene (1) in addition to the known 7α-acetoxy-12-hydroxy- 11,14-dioxoabieta-8,12-diene (7α-acetoxyroyleanone) (2). Structures of 1 and 2 were established by spectroscopic means. The cytotoxic activity against five cancer cell lines was evaluated. Compounds 1 and 2 displayed considerable cytotoxic activity against several cancer cell lines with IC 50 values in the range of 3.1 to 11.5 μg/ml and 0.9 to 7.6 μg/ml, respectively. The cytotoxic activity seemed to be related to inhibition of DNA synthesis, as revealed by the reduction of 5-bromo-2′-deoxyuridine incorporation and induction of apoptosis, as indicated by the acridine orange/ethidium bromide assay and morphological changes after 24 h of incubation in leukemic cells.

A new flavanone, 7-hydroxy-5,6-dimethoxyflavanone (1), together with three other flavonoids, didymocarpin (2), 2′,4′-dihydroxy-5′,6′-dimethoxychalcone (3), and isodidymocarpin (4), had been isolated from the methanol extract of the tree bark of Cryptocarya costata. The structures of these compounds were determined based on spectral evidence, including UV, IR, 1-D and 2-D NMR, and mass spectra. Cytotoxic properties of compounds 1-4 were evaluated against murine leukemia P-388 cells. The chalcones 3 and 4 were found to have substantial cytotoxicity with IC 50 of 5.7 and 11.1 μᴍ, respectively.

Bioassay-guided fractionation of a Satureja parvifolia MeOH extract led to the isolation of eriodictyol, luteolin and ursolic and oleanolic acids as its active components against Plasmodium falciparum K1. This is the first time these compounds are reported as constituents of S. parvifolia. Ursolic acid showed an IC 50 of 4.9 μg/ml, luteolin 6.4 μg/ml, oleanolic acid 9.3 μg/ml and eriodictyol 17.2 μg/ml. Antiplasmodial activity of eriodictyol and luteolin is reported here for the first time. Besides, the four compounds showed activity against P. falciparum 3D7 strain and Trypanosoma brucei rhodesiense. Eriodictyol showed moderate activity on all the parasites but was the most selective compound as a result of its rather low cytotoxicity (IC 50 174.2 μg/ml) on the mammalian KB cell line.

A new tetranortriterpene 3α-acetoxy-24,25,26,27-tetranortirucalla-7-ene-23(21)-lactone (3), and eleven other compounds were isolated from the twigs of Amoora dasyclada. The structure of compound 3 was identified on the basis of spectroscopic data, and the bioactive experiments of 1 and 3-5 against AGZY 83-a (human lung cancer cells) and SMMC-7721 (human liver cancer cells) are documented. Among them, compound 5 exhibited a strong activity against SMMC-7721.

The phytochemistry of the genus Piper (Piperaceae) has been widely studied due to the biological properties of amides from these plants. In this work, we have synthesized and evaluated the toxic effect of 11 amides against the fall armyworm Spodoptera frugiperda larvae. The naturally occurring piperine was also evaluated. The most active amide was N- [3-(3′,4′-methylenedioxyphenyl)-2-(E)-propenoyl]piperidine with a LD 50 of 1.07 μg mg −1 larvae. This amide was also evaluated by ingestion.

We characterized a newly isolated bacterium, designated as IR1, with respect to its ability to degrade polycyclic aromatic hydrocarbons (PAHs) and to produce biosurfactants. Isolated IR1 was identified as Pseudomonas putida by analysis of 16S rRNA sequences (99.6% homology). It was capable of utilizing two-, three- and four-ring PAHs but not hexadecane and octadecane as a sole carbon and energy source. PCR and DNA hybridization studies showed that enzymes involved in PAH metabolism were related to the naphthalene dioxygenase pathway. Observation of both tensio-active and emulsifying activities indicated that biosurfactants were produced by IR1 during growth on both water miscible and immiscible substrates. The biosurfactants lowered the surface tension of medium from 54.9 dN cm −1 to 35.4 dN cm −1 and formed a stable and compact emulsion with an emulsifying activity of 74% with diesel oil, when grown on dextrose. These findings indicate that this isolate may be useful for bioremediation of sites contaminated with aromatic hydrocarbons.

Soils at agrochemical dealer sites often are contaminated with pesticide residues from decades of accidental and incidental spillage. We have determined that prairie grasses native to the Midwestern U.S. are suitable for phytoremediation because they are tolerant of most herbicides and of climatic extremes, such as heat, cold, drought, and flooding. A mixed stand of big bluestem, switch grass, and yellow indiangrass develops a rhizosphere with microflora that can readily detoxify pesticide residues. Specific atrazine-degrading bacteria or the free enzyme atrazine chlorohydrolase also can enhance the rate of biotransformation of atrazine in soil. Metolachlor degradation can be accelerated significantly by the prairie grass/rhizosphere effect. Several grasses used in filter strips have also been evaluated for their pesticidedegradation capabilities. The prairie grasses also have been demonstrated to reduce the rates of leaching of pesticides through intact soil columns, since less water leaches out of vegetated soil columns compared to non-vegetated soil columns. The evaluation of the degree of success of remediation has relied heavily on chemical residue analysis, but recent studies on biological endpoints have shown promise for providing more ecologically relevant indications of the potential exposure of organisms to pesticides in the soil. Earthworm 8-day bioaccumulation assays and root growth assays have shown the value of assessing the bioavailability of the residues. Mass balance experiments have utilized radiolabeled atrazine and metolachlor to ascertain the complete metabolism and binding profile of those two pesticides in phytoremediation studies.

The protein mobilization from attached and detached seeds of Vicia faba L. cv. Eresen 87 (Fabaceae) was investigated. While the total soluble protein content decreased, the free amino acid content increased during the 7 days germination period. Among the three proteolytic enzymes, only endopeptidase activity was found to be affected by the removal of the embryonic axis. Leucine aminopeptidase activity was high at the beginning, then it decreased; carboxypeptidase activity reached the highest value at day 5. In order to examine the effects of plant growth regulators on detached cotyledons incubated with plant growth regulators [10 −4 ᴍ benzyladenine (BA), gibberellic acid (GA 3 ), indole acetic acid (IAA) and 10 −5 ᴍ abscisic acid (ABA)], only benzyladenine was found promotive on protein mobilization. Our results suggest that the removal of the embryonic axis in seeds of Vicia faba L. cv. Eresen 87 decreases protein mobilization and endopeptidase activity.

6α,7β-Dihydroxyvouacapan-17β-oic acid (1) was isolated from Pterodon polygalaeflorus Benth. Modification of 1 yielded 6α-hydroxyvouacapan-7β,17β-lactone (2) and then 6-oxovouacapan- 7β,17β-lactone (3). Photosynthesis inhibition by 3 was evaluated in spinach chloroplasts. The uncoupled non-cyclic electron transport rate and ATP synthesis were inhibited by 3, which behaved as a Hill reaction inhibitor. Furthermore, 3 acted as an uncoupler because it enhanced the basal and phosphorylating electron transport rate on thylakoids. This last property of 3 was corroborated when it was observed that it enhances the Mg 2+ -ATPase activity. In contrast, 3 did not affect photosystem I (PSI) activity. Analysis of the partial photosystem II (PSII) reactions from water to DCPIPox and water to silicomolybdate allowed to locate the inhibition sites at the redox components of PSII. The OJIP test of the chlorophyll a fluorescence transient confirmed that the inhibition sites were 1.) the oxygen-evolving complex (OEC) and 2.) by the formation of silent centers in the non-QA reducing centers.

The stability of PSII in leaves of the resurrection plant Haberlea rhodopensis to high temperature and high light intensities was studied by means of chlorophyll fluorescence measurements. The photochemical efficiency of PSII in well-hydrated Haberlea leaves was not significantly influenced by temperatures up to 40 °C. F 0 reached a maximum at 50 °C, which is connected with blocking of electron transport in reaction center II. The intrinsic efficiency of PSII photochemistry, monitored as F v /F m was less vulnerable to heat stress than the quantum yield of PSII electron transport under illumination (ΦPSII). The reduction of ΦPSII values was mainly due to a decrease in the proportion of open PSII centers (qP). Haberlea rhodopensis was very sensitive to photoinhibition. The light intensity of 120 μmol m −2 s −1 sharply decreased the quantum yield of PSII photochemistry and it was almost fully inhibited at 350 μmol m −2 s −1 . As could be expected decreased photochemical efficiency of PSII was accompanied by increased proportion of thermal energy dissipation, which is considered as a protective effect regulating the light energy distribution in PSII. When differentiating between the three components of qN it was evident that the energy-dependent quenching, qE, was prevailing over photoinhibitory quenching, qI, and the quenching related to state 1-state 2 transitions, qT, at all light intensities at 25 °C. However, the qE values declined with increasing temperature and light intensities. The qI was higher than qE at 40 °C and it was the major part of qN at 45 °C, indicating a progressing photoinhibition of the photosynthetic apparatus.

The influence of elicitation on rosmarinic acid biosynthesis by Lavandula vera MM cell suspension culture was investigated using vanadyl sulfate as an abiotic elicitor. It was established that 12 h after treatment with 25 mg/l vanadyl sulfate the rosmarinic acid production was increased up to 3.92 g/l (2.8 times higher compared to the control cultivation). No significant amounts of rosmarinic acid were detected in the culture medium in comparison with its intracellular content. However, it was observed that the extracellular content of rosmarinic acid is 3.3 times higher compared to the control variant (4 h after treatment at elicitor concentration 25 mg/l).

The effects of salicylic acid (SA) on the accumulation of dehydrins in leaves of Tibetan hulless barley seedlings under water stress were investigated. The results indicated that SA decreased the levels of the four dehydrin-like proteins induced by water stress. The concentrations of these dehydrin-like proteins increased under water stress. However, their levels in SA-pretreated seedlings were always lower than in those receiving only water stress. Our results also indicated that the levels of dehydrin-like proteins decreased as the SA concentration increased. In SA-pretreated seedlings, electrolyte leakage, MDA and H 2 O 2 content were rather higher than in seedlings receiving only water stress. By these results, we suggest that lower levels of dehydrin-like proteins in seedlings with SA treatment may be due to the greater accumulation of H 2 O 2 induced by SA, which causes more oxidative injury under water stress.

An efficient procedure has been developed for callus induction and cell suspension cultures of C. saxicola for the first time. Explant selection was carried out among leaf, stem and root to select a suitable type of explants capable of higher callus formation. Leaf explants thus selected showed maximum response to callus induction (67.1%). Modified B 5 medium supplemented with 0.5 mg l −1 2,4-D plus 2 mg l −1 BA was the most favorable medium for callus formation with the highest induction rate (94.8%) and greatest fresh weight of callus (1.7 g per explant). Cell suspension cultures were established by transferring 2-8 g fresh callus to 80 ml liquid B5 medium. An inoculum size of 8 g produced the greatest biomass accumulation, dehydrocavidine and berberine productions, which was 13.1 g l −1 , 8.0 mg l −1 and 4.1 mg l −1 , respectively. In response to various sucrose concentrations from 10 g l −1 to 80 g l −1 , cultures with 60 g sucrose l −1 not only produced the highest dry biomass (18.5 g l −1 ) but also the highest formation of dehydrocavidine (11.6 mg l −1 ) and berberine (7.6 mg l −1 ). These prepared cell suspension cultures provided a useful material for further regulation of alkaloid biosynthesis and for enhanced production of valuable alkaloids on a large scale.

Contents of three 1,4-benzoxazin-3-ones in tissue samples from different parts (young leaf, second leaf, old leaf, stem and root) of young maize plants of 4-leaves stage, fed by the third instar larvae of the Asian corn borer, Ostrinia furnacalis (Guenée), were analyzed by highperformance liquid chromatography-mass spectroscopy (HPLC-MS). Samples were taken immediately (set A) or 48 h (set B) after larvae had fed on the second leaf for 48 h. The three 1,4-benzoxazin-3-ones investigated in our experiments were 2,4-dihydroxy-7-methoxy- 1,4(2H)-benzoxazin-3-one (DIMBOA), 2,4-dihydroxy-1,4(2H)-benzoxazin-3-one (DIBOA) and 2-hydroxy-7-methoxy-1,4(2H)-benzoxazin-3-one (HMBOA). In samples of set A, the levels of DIMBOA and HMBOA were significantly lifted in the old leaf (L3) and young leaf (L1), respectively, while amounts of these two chemicals in other plant parts were not significantly different between larvae-fed plants and intact plants. Concentrations of DIBOA in each plant part remained unchanged. In samples of set B, no concentration differences for any of these three 1,4-benzoxazin-3-ones between larvae-fed plants and controls were observed in any plant part. The feeding of the Asian corn borer seems to have limited effects on induction of these three 1,4-benzoxazin-3-ones in young maize plants of the variety investigated.

By X-ray structure analysis it could be shown that from the solution equilibrium of pyochelin I and II, differing in the stereochemistry at C-2″ (1a and 1b), crystals of the Fe 3+ complex of the steroisomer 1a are formed with a 1:1 metal-to-ligand ratio. Ligand sites are the carboxylate and the phenolate anions and the two nitrogen atoms. Two equivalent ferri-pyochelin moieties are held together by a hydroxy and an acetate unit which satisfy the remaining two coordination sites of Fe 3+ .

Cell suspension cultures are now recognized as important model materials for plant bioscience and biotechnology. Very few studies of metabolic comparisons between cell cultures and original plants have been reported, even though the biological identity of cultured cells with the normally grown plant is of great importance. In this study, a comparison of the metabolome for primary metabolites extracted from the leaves of Arabidopsis thaliana and cultured cells from an Arabidopsis suspension culture (cell line T87) was performed. The results suggest that although cell suspension cultures and Arabidopsis leaves showed similarities in the common primary metabolite profile, nonetheless, moderate differences in quantitative profile were revealed.

We have confirmed that dissociation of the dimeric SOD molecule into a monomeric one can be readily detected in solution by the use of capillary electrophoresis (CE), which is based on the fact that the peak height in the CE profile is highly dependent on the aggregation conditions of the protein molecule. Based on this fact, it has become apparent that the hydrogen peroxide molecule induces the dissociation of the dimeric structure of SOD, and this should give reasonable explanation for the inactivation of SOD by hydrogen peroxide. Our results may give a convenient way for the early detection of the amyotrophic lateral sclerosis in patients, because we can estimate whether the SOD molecule is of a rigid or loosed dimeric structure by the use of this technique. The loosed one has been assumed to exhibit inherent toxicity of the copper center, so-called “gain-of-function” of the mutant SOD.

Gas chromatography and mass spectrometry analyses of crude sex pheromone gland extracts revealed that virgin Synanthedon tipuliformis (Clerck), currant borer (Lepidoptera: Sesiidae) females, produced 6 compounds, structurally related to sex pheromone components of clearwing moths. By comparison of retention times and mass spectra of natural products with corresponding properties of synthetic standards, these compounds were identified as: (2E,13Z)-octadeca-2,13-dien-1-yl acetate (E2,Z13-18:OAc), (3E,13Z)-octadeca-3,13-dien-1-yl acetate (E3,Z13-18:OAc), (13Z)-octadec-13-en-1-yl acetate (Z13-18:OAc), (2E,13Z)-octadeca- 2,13-dien-1-ol (E2,Z13-18:OH), (13Z)-octadec-13-en-1-ol (Z13-18:OH) and octadecan- 1-ol (18:OH) in the ratio 100:0.7:2.7:3.2:traces:traces. The first 3 compounds were previously known to occur in the sex pheromone gland extracts of currant borers, while the last 3 chemicals are now reported for the first time. Trapping tests carried out in the black currant field revealed that E2,Z13-18:OAc, when tested separately, attracted S. tipuliformis males, while addition of E3,Z13-18:OAc to the main component increased the effectiveness of E2,Z13-18:OAc over seven times. The attractiveness of 6 component lures did not differ significantly from the one of the binary mixture, confirming that E2,Z13-18:OAc and E3,Z13- 18:OAc in the ratio100:0.7 are essential sex pheromone components of S. tipuliformis. Trapping tests carried out at the dwelling place of Synanthedon scoliaeformis (Borkhausen) (Lepidoptera: Sesiidae) revealed that, in addition to intraspecific synergistic effect, E3,Z13-18:OAc increased the specificity of the pheromone signal of S. tipuliformis, acting by intraspecific mode as an attraction antagonist against S. scoliaeformis males. By this way, it ensured the specificity of the sex attraction signal of the currant borer. Consequently, both compounds E2,Z13-18:OAc and E3,Z13-18:OAc have to be present in pheromone formulations used for monitoring and/or control of S. tipuliformis to avoid effecting non-target species. Other compounds identified from the sex pheromone gland of S. tipuliformis did not show any significant interspecific activity for males of S. scoliaeformis, however, they provide a basis to achieve specificity of a pheromone signal of S. tipuliformis and could act as attraction antagonists against other clearwing moth species which, like S. tipuliformis, employ E2,Z13- 18:OAc as their sex pheromone component.

The role of vitamin A was evaluated for its chemotherapeutic and chemoprophylactic action against Acanthocheilonema viteae infection in Mastomys coucha. Vitamin A was administered for 10 days, five days before infection and five days post infection. On day 0 experimental animals as well as controls were infected with L3, the infective stage. Establishment of the worms revealed significantly less percentage of worm recovery over untreated controls. Cell-mediated response was found to be the cause of this reduction in worm recovery, whereas humoral response was not significant as lgG, lgA and lgM titres were low.

Kinetics and the mechanism of total in vitro hydrolyses (i.e. up to the exhaustion of substrate) of acetylcholine and acetylthiocholine by acetylcholinesterase and butyrylcholinesterase were studied in vitro in a batch reactor at 25 °C, pH 8 and ionic strength of 0.11 ᴍ. Every hydrolysis was monitored by 2 - 3 independent analytical methods. All studied types of enzymatic hydrolyses fulfilled the Michaelis - Menten reaction scheme with the irreversible second step. A table of obtained average values of rate constants and estimations of initial molar enzyme concentrations, and discussion of the results are presented.

An indirect format enzyme-linked immuno-sorbent assay (ELISA) on graphite rods (Ø 0.8 mm x 20 mm) and, for comparison, on microtiter plates has been developed against terbuthylazine. For this purpose, a series of 2-aminoalkyl-4-chloro-6-terbuthyl-s-triazine-2,6- diamine ELISA haptens with alkyl spacer lengths of 2, 4, 6, and 8 CH 2 groups has been synthesized. The graphite rods or the microtiter plates were covered by a polymerized glutaraldehyde network, and the ELISA haptens have been coupled by imino coupling to the free aldehyde groups of that network. ε-Aminocaproic acid has been used as an agent to block unspecific binding sites. The ELISA was run in a competitive mode, where the immobilized ELISA hapten and the solute terbuthylazine as a target analyte compete for the solute antibody.

Effects of N-dodecyl-N,N-dimethyl-N-benzylammonium halides (DBeAX) on thermotropic phase behavior of phosphatidylcholine/cholesterol bilayers as well as on 1 H NMR spectra were studied. The surfactants were added either to the water phase or directly to the lipid phase (a mixed film was formed). The benzyl group, opposite to liposomes without cholesterol, is not incorporated into the bilayer in the gel state but only in the liquid state. All the halides DBeAX (particularly the chloride DBeAC) showed greater ability to destabilize the membrane structure in the presence than in the absence of cholesterol. The interaction of DBeAX with DPPC/cholesterol bilayers and subsequent changes in the phospholipid bilayer organization depended on the kind of counterion. The strongest effects were observed for chloride (most electronegative ion) and for iodide (largest ion). The effects of chloride and bromide on phase transition and 1 H NMR spectra in the presence and absence of cholesterol were opposite. This is discussed in terms of the influence of counterions on the pair cholesterol-DPPC interactions.