Volume 59, Issue 7-8

Leaf glands of Ceanothus species excrete a lipophilic material that contains a variety of flavonoids. Most of these are aglycones, but some glycosides were also observed. Seven out of eight species exhibit flavonols, whereas flavones are excreted by only one species. Four species produce flavanones and dihydroflavonols; one excretes a remarkable quantity of flavonol glycosides. The exudate flavonoids thus form different patterns that might be characteristic for different Ceanothus species.

The oils obtained by hydrodistillation and steam distillation of the aerial parts of Stachys persica Gmel. and Stachys byzantina C. Koch grown in Iran were analyzed by GC/MS. The essential oil obtained by hydrodistillation of the aerial parts of S. persica was characterized by a high amount of non-terpenoid components of which methyllinoleate (27.7%), hexadecanoic acid (9.8%) and 6,10,14-trimethyl-2-pentadecanone (9.2%) were the major constituents, whereas the steam distilled oil of the plant contained hexadecanoic acid (27.2%), carvacrol (9.4%) and eugenol (5.2%). Both hydrodistilled and steam distilled essential oils of the aerial parts of S. byzantina were rich in sesquiterpenes such as α-copaene (16.6% and 10.4%), spathulenol (16.1% and 18.5%) and β-caryophyllene (14.3% and 13.5%), respectively.

The immunomodulatory bioassay-guided fractionation of the methanolic extract of henna (Lawsonia inermis L.; syn. Lawsonia alba L.) leaves resulted in the isolation of seven compounds; three have been isolated for the first time from the genus, namely p-coumaric acid, 2-methoxy-3-methyl-1,4-naphthoquinone and apiin, along with the previously isolated compounds: lawsone, apigenin, luteolin, and cosmosiin. Structural elucidation of the isolated compounds was based upon their physical, chemical as well as spectroscopic characters. Their immuomodulatory profile was studied using an in vitro immunoassay, the lymphocyte transformation assay. The ABTS [2,2′-azino-bis (3-ethyl benzthiazoline-6-sulfonic acid)], free radical scavenging assay depicted that all isolated compounds exhibited antioxidant activity comparable to that of ascorbic acid.

A free radical scavenging activity guided fractionation of the polar extract from roots of Limonium brasiliense (Plumbaginaceae) led to the isolation of five active compounds including: myricetin 3-O-α-rhamnopyranoside (1), (-)-epigallocatechin 3-O-gallate (2), (-)-epigallocatechin (3), (+)-gallocatechin (4) and gallic acid (5). These and other chemical constituents are reported for the first time for this species. The characterization of these compounds was achieved by spectroscopic methods ( 1 H NMR, 13 C NMR and UV).

In vitro antioxidant activities of three sweet dihydrochalcone glucosides from the leaves of Lithocarpus pachyphyllus (Kurz) Rehd. (Fagaceae), trilobatin 2″-acetate (1), phloridzin (2) and trilobatin (3), were investigated. The IC 50 (50% inhibitory concentration) values for compounds 1-3 of lipid peroxidation in rat liver homogenate were 261, 28, 88 μm, respectively. Compounds 1-3 increased superoxide dismutase (SOD) activity with EC 50 (50% effective concentration) values of 575, 167, 128 μm, and glutathione peroxidase (GSH-Px) activity with EC 50 values of 717, 347, 129 μm, respectively, and showed only weak DPPH (1,1- diphenyl-2-picrylhydrazyl) radical scavenging activity

Looking for new plant sources of immunomodulating agents polysaccharide-rich fractions (PS) from Menyanthes trifoliata L. (Menyanthaceae) have been isolated. The herb of Menyanthes trifoliata L. was sequentially extracted with water, 0.1 м NaOH, 8% CH 3 COOH, and 1 м NaOH. After dialysis and resolution on Biogel P-10 four homogenic (B-4, B-5, C-4, D-5) and two nonhomogenic (A-3 and D-4) PS were isolated. About 0.5% of PS over 3500 Da were found in the dry plant material. They were characterized through chemical analysis, NMR and vibrational spectroscopy. Speciation analysis of chosen metal/metaloid elements was performed and an exceptionally high concentration of Se was found in PS of a pure water extract (A-3). The biological tests on the immunomodulating influence with human blood-derived lymphocytes and granulocytes revealed that two fractions, B-4 and B-5, were strong stimulators of immune cells, whereas fractions D-5 and A-3 were found as potent suppressive and anti-inflammatory agents. The applied isolation procedures led to the separation of active compounds into stimulatory and inhibitory fractions.

Three lignan derivatives, (-)-taxiresinol (1), (-)-3′-demethylisolariciresinol-9′-hydroxyisopropylether (2) and (-)-3-demethylisolariciresinol (3), previously isolated from the heartwood of Taxus baccata L. (Taxaceae) were investigated for cytotoxicity against the Oncology Cell Line Panel (breast, colon, ovary, prostate, lung and a normal adult bovine aortic endothelial cell line) as well as for antimicrobial activities. Besides, a chloroform-soluble portion of the ethanol extract from the heartwood of T. baccata was also tested for antimicrobial activities. Compounds 1 - 3 did not demonstrate much cytotoxic potency according to the reference drug etoposide. With the exception of compound 2, the other two lignans (1 and 3) and the chloroform extract were shown to possess antifungal activity, whereas only the chloroform extract exhibited antibacterial activity against Pseudomonas aeruginosa

A new aurone 1 and two known substances, aurantiamide acetate (2) and tiliroside (3), were isolated from the ethanolic extract of Gomphrena agrestis. The structural determination of 1 was based on spectroscopic and spectrometric data. The substance was defined as (E)-3′- O-β-ᴅ-glucopyranosyl-4,5,6,4′-tetrahydroxy-7,2′-dimethoxyaurone. Biological activity of the ethanolic crude extract and isolated compounds against bacteria, fungi and Leishmania amazonensis amastigotes was evaluated. This appears to be the first report documenting aurone and aurantiamide compounds in the Amaranthaceae family. In the evaluation of biological activity the ethanolic extract of G. agrestis and compounds 1, 2, and 3 were shown to be active mainly against Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa.

The chloroform extract of Ferula persica var. persica roots was found to inhibit red pigment production of Serratia marcescens. A bioguided fractionation study by preparative thin layer chromatography (PTLC) detected a fraction (R f = 0.71, petroleum ether/EtOAc, 2:1 v/v), which was effective on depigmentation of Serratia marcescens. Using conventional spectroscopy methods, the active fraction was identified as umbelliprenin. Neither the chloroform extract nor the isolated umbelliprenin fraction showed any antibacterial activity against the test strain at a certain concentration. In contrast, they exhibited depigmentation zones on culture plates.

Five phenolic compounds, 4-hydroxybenzoic acid methyl ester (1), vanillic acid methyl ester (2), 4-hydroxy benzaldehyde (3), 4-hydroxybenzoic acid (4) and ferulic acid (5), and four flavonoids, 5,5′-dihydroxy-4′,6,7-trimethoxyflavanone (6), luteolin (7), vitexicarpin (8) and artemetin (9), were isolated from fruits and leaves of Vitex rotundifolia L. The biological activities of these nine compounds have been examined using a bioassay with lettuce seedlings.

The potential biological activity of binary mixtures of some new organophosphorous compounds, aminoalkane- and aminofluorenephosphonates, with (N-phosphonomethyl)-glycine (glyphosate, PMG) was studied. The inhibition of growth of wheat (Triticum aestivum) induced by individual compounds and their equimolar mixtures with PMG was a measure of that activity. The experiments were expected to show if the new compounds exhibited good biological activity to be used for agrochemical applications and if this activity can be improved when they are used in mixtures with glyphosate which is the active component of the well-known herbicide Roundup. The results obtained show that aminofluorenephosphonates inhibited wheat growth when used in micromolar concentrations. Thus, their efficiency can be compared to that of PMG. The efficiency of aminoalkanephosphonates was one order of magnitude weaker. The measure of the efficiency was the effective concentration inhibiting wheat growth by 50% (EC 50 ). The most demanded interaction, i.e., a synergistic was observed for only one of binary mixtures of the compounds studied with PMG. Mostly they showed antagonistic or strong antagonistic interactions. Some of them were of the additive type. Such results exclude the possibility of potential use of all the compounds studied in binary mixtures with phosphonomethylglycine, especially as the mentioned synergistic interaction found was rather weak. The influence of structural features of aminophosphonates on the results obtained is discussed.

Pisolithus tinctorius (Basidiomycete) is an ectomicorrhizal fungus found in the roots and soil surrounding of many species of eucalyptus and pine trees. The present work verified the cytotoxic potential of pisosterol, a triterpene isolated from P. tinctorius collected in the Northeast region of Brazil, on three different animal cell models: mouse erythrocytes, sea urchin embryos and tumor cells. Pisosterol lacked activity on mouse erythrocytes as well as on the development of sea urchin eggs, but strongly inhibited the growth of all seven tumor cell lines tested, especially the leukemia and melanoma cells (IC 50 of 1.55, 1.84 and 1.65 μg/ ml for CEM, HL-60 and B16, respectively). The results found for pisosterol were compared with those of doxorubicin and etoposide

The aim of this work was to study the antiproliferative effect of a tincture from fruits of Angelica archangelica and the active components using the human pancreas cancer cell line PANC-1 as a model. Significant dose-dependent antiproliferative activity was observed in the tincture with an EC 50 value of 28.6 μg/ml. Strong antiproliferative activity resulted from the two most abundant furanocoumarins in the tincture, imperatorin and xanthotoxin. The contribution of terpenes to this activity was insignificant. Imperatorin and xanthotoxin proved to be highly antiproliferative, with EC 50 values of 2.7 μg/ml and 3.7 μg/ml, respectively, equivalent to 10 and 17 μm. The results indicate that furanocoumarins account for most of the antiproliferative activity of the tincture.

The latex of Croton draco, its extracts and several latex components have been investigated for their influence on both classical (CP) and alternative (AP) activation pathways of the complement system using a hemolytic assay. The best inhibition was found for the classical pathway. The latex, ethyl acetate and ethyl ether extracts exhibited extremely high inhibition on the CP (94, 90 and 77%, respectively) at a concentration of 1 mg/ml. The flavonoid myricitrin, the alkaloid taspine and the cyclopeptides P1 and P2 showed high inhibition on CP (83, 91, 78 and 63%, respectively) at a concentration of 0.9 mм.

In this paper, a headspace solid-phase microextraction (HS-SPME) method was applied to analyse the volatile compounds in a traditional Chinese medicine (TCM), Rhioxma Curcumae Aeruginosae. SPME parameters such as fibers, extraction temperature, extraction time and desorption time were investigated. Thirty-five volatile compounds were separated and identified. Relative standard deviations (RSDs) were less than 8.4%, showing that the method has a good reproducibility. The volatile constituents were also analyzed by steam distillation (SD) and thirty-seven compounds were identified. The similar results obtained by the two methods showed that SPME is a good alternative for the analysis of volatile constituents in Rhioxma Curcumae Aeruginosae samples and it is a relatively simple, rapid and solvent-free method.

A single cell of the green paramecia (Paramecium bursaria) harbors several hundreds of endo-symbiotic Chlorella-like algae in its cytoplasm. Removal of algae from the host organism and re-association of ex-symbiotic host paramecia with ex-symbiotic algae can be experimentally demonstrated in the laboratory. However, the mechanism precisely governing the alga-protozoan association is not fully understood, and the origin of symbiosis in the evolutionary view has not been given. Here, we propose the possible biochemical models (models 1 and 2) explaining the co-evolution between Paramecium species and algal symbionts by pointing out that algal photosynthesis in the host paramecia plays a dual role providing the energy source and the risk of oxidative damage to the host. Model 1 lays stress on the correlation between the (re)greening ability of the paramecia and the tolerance to oxidative stress whereas model 2 emphasizes the cause of evolutionary selection leading to the emergence of Paramecium species tolerant against reactive oxygen species.

The responses of young plants of diploid and tetraploid Matricaria chamomilla cultivars to abiotic stress were studied. The course of quantitative changes of main leaf secondary metabolites was evaluated within an interval from 6 h before to 54 h after spraying the leaf rosettes with aqueous CuCl 2 solution. The content of herniarin in the treated plants rose approximately 3 times, simultaneously with a decline of its precursor (Z)- and (E)-2-β-ᴅ-glucopyranosyloxy- 4-methoxycinnamic acid. The highest amounts of umbelliferone in stressed plants exceeded 9 times and 20 times those observed in control plants of the tetraploid and diploid cultivar, respectively. Due to stress the concentration of ene-yne-dicycloether in leaves decreased by more than 40%. The pattern of quantity changes of the examined compounds in tetraploid and diploid plants was similar.

The first elongation step to form very-long-chain fatty acids (VLCFAs) is catalyzed by the VLCFA-synthase. CoA-activated fatty acids react with malonyl-CoA to condense a C2-unit. As shown with recombinant enzyme this reaction is specifically inhibited by chloroacetamide herbicides. The inhibition is alleviated when the inhibitor (e.g. metazachlor) is incubated together with adequate concentrations of the substrate (e.g. oleoyl-CoA). Malonyl-CoA has no influence. However, once a chloroacetamide has been tightly bound to the synthase after an appropriate time it cannot be displaced anymore by the substrate. In contrast, oleoyl- CoA, is easily removed from the synthase by metazachlor. The irreversible binding of the chloroacetamides and their competition with the substrate explains the very low half-inhibition values of 10 -8 м and below. Chiral chloroacetamides like metolachlor or dimethenamid give identical results. However, only the (S)-enantiomers are active.

Agrobacteria mediated Coleus blumei tumour tissues were cultured in vitro on MS medium. Sixteen diversified transformed callus cultures were maintained for several years in the absence of plant growth regulators and antibiotics without affecting the growth rate. Rosmarinic acid was detected spectrophotometrically in all tissue lines but in different quantities. The highest rosmarinic acid accumulation detected was 11% of dry tissue mass. The relation between culture growth and rosmarinic acid production was investigated in three callus lines. The lines showed different rosmarinic acid accumulation in relation to their growth rate; it was either parallel or inversely related to the tissue growth. The effects of certain medium constituents on the callus growth and rosmarinic acid accumulation were examined in four tumour cell lines. Addition of 4% or 5% sucrose stimulated rosmarinic acid synthesis and decreased callus growth. Nitrogen reduction to one half or one quarter of initial concentration did not affect rosmarinic acid synthesis and decreased callus growth in three lines, while it increased rosmarinic acid accumulation and callus growth in one line. Addition of 0.1 mg/l Phe stimulated rosmarinic acid production in two lines but had little effect on the rosmarinic acid level in others. Rosmarinic acid production was significantly improved on modified macronutrients, where the Ac2 line produced 16.5 mg of rosmarinic acid per tube (0.2 g of dry wt) after being in culture for 35 days.

Two full-length cDNAs (TCH1 and TCH2) were obtained from a cDNA library of Taxus chinensis mainly by the single specific-primer PCR (SSP-PCR) method. Compared with other reported enzymes from Taxus species, the deduced amino acid sequences of TCH1 and TCH2 exhibit significant homologies to hydroxylases that are involved in taxol biosynthesis. These findings imply that the two new genes are closely related to the biosynthesis of taxol/taxoids.

Using several consensus sequences for the 106 amino acid residue α-spectrin repeat segment as probes we searched animal sequence databases using the BLAST program in order to find proteins revealing limited, but significant similarity to spectrin. Among many spectrins and proteins from the spectrin-α-actinin-dystrophin family as well as sequences showing a rather high degree of similarity in very short stretches, we found seven homologous animal sequences of low overall similarity to spectrin but showing the presence of one or more spectrin-repeat motifs. The homology relationship of these sequences to α-spectrin was further analysed using the SEMIHOM program. Depending on the probe, these segments showed the presence of 6 to 26 identical amino acid residues and a variable number of semihomologous residues. Moreover, we found six protein sequences, which contained a sequence fragment sharing the SH3 (sarc homology region 3) domain homology of 42-59% similarity. Our data indicate the occurrence of motifs of significant homology to α-spectrin repeat segments among animal proteins, which are not classical members of the spectrin-α- actinin-dystrophin family. This might indicate that these segments together with the SH3 domain motif are conserved in proteins which possibly at the early stage of evolution were close cognates of spectrin-α-actinin-dystrophin progenitors but then evolved separately.

Random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) markers were used to study the genomic relationship among 11 members of Indian Rhizophoraceae represented by nine true mangroves and two non-mangrove species. The AFLP and RAPD bands were scored and analyzed for genetic similarities and cluster analysis was done which separated the 11 species studied into two main groups, the true mangroves and the non-mangroves. The polymorphism observed for these markers showed a high degree of genetic diversity among the constituent taxa of the family. The phylogenetic relationship inferred from molecular marker systems supported the traditional taxonomic classification of the family Rhizophoraceae based on morphological characters at the levels of tribe, phylogeny and delimitation of genera and species, except the intra-generic classification of the genus Bruguiera and the placement of Rhizophora in the family Rhizophoraceae.

Secretion products from the opisthosomal defense gland of south east Asian whip scorpions were identified for the first time by gas-chromatography and mass-spectrometry. Specimens of the genera Hypoctonus, Typopeltis and Ginosigma were tested. While some ingredients are present in large concentrations, others are possibly only side products and may be synthesized more incidentally. For this reason no important functional role is attributed to them. There are considerable individual differences concerning the concentrations of various ingredients. While the secretion products of most species of the genus Typopeltis - similar to Mastigoproctus - are characterized by acetic and octanoic acid in large concentrations, the secretion product of Hypoctonus siamensis provides octanoic acid only in a very low concentration but it is characterized by hexyl acetate.

Secretions of three different glands (mandibular gland, labial gland, and Dufour’s gland) of virgin queens of five bumblebee species (Bombus lucorum, B. lapidarius, B. hypnorum, B. pascuorum, and B. terrestris) were analysed. Around 200 compounds were identified in the secretions. The compositions of the secretions of labial and mandibular glands were speciesspecific. Dufour’s gland of all species produced mainly hydrocarbons, both saturated and unsaturated, the proportions of which differed quantitatively between the species studied

Proteases mediate important crucial functions in parasitic diseases, and their characterization contributes to the understanding of host-parasite interaction. A serine protease was purified about 43-fold with a total recovery of 60% from a detergent-soluble extract of promastigotes of Leishmania amazonensis. The purification procedures included aprotinin-agarose affinity chromatography and gel filtration high performance liquid chromatography. The molecular mass of active enzyme was 110 kDa by native gel filtration HPLC and by SDSPAGE gelatin under non-reducing conditions. Under conditions of reduction using SDSPAGE gelatin analyses the activity of enzyme was observed in two proteins of 60 and 45 kDa, suggesting that the enzyme may be considered as a dimer. The Leishmania protease was not glycosylated, and its isoelectric point (pI) was around 4.8. The maximal protease activity was at pH 7.0 and 28 °C, using α-N-ᵨ-tosyl-ʟ-arginyl-methyl ester (ʟ-TAME) as substrate. Assays of thermal stability indicated that this enzyme was totally denatured after pre-treatment at 42 °C for 12 min and preserved only 20% of its activity after pre-treatment at 37 °C for 24 h, in the absence of substrate. Hemoglobin, bovine serum albumin (BSA), ovalbumin and gelatin were hydrolyzed by Leishmania protease. Inhibition studies indicated that the enzyme belonged to a serine protease class because of a significant impediment by serine protease inhibitors such as benzamidine, aprotinin, and antipain. The activity of the present serine protease is negatively modulated by calcium and zinc and positively modulated by manganese ions. This is the first study that reports the purification of a protease from a detergent-soluble extract of Leishmania species.

The inhibitory potency of four alkylphospholipids: rac-1-O-phosphocholine-2-hydroxy-octadecane (rac-2-OH), rac-1-O-phosphocholine-2-O-acetyl-octadecane (rac-2-O-acetyl), rac-1- O-phosphocholine-2-amino-octadecane (rac-2-NH 2 ) and rac-1-O-phosphocholine-2-N-acetyloctadecane (rac-2-N-acetyl), on the cytochrome P450-dependent monooxygenase activity has been evaluated. The IC 50 values of the alkylphosphocholines with 7-ethoxycoumarin as substrate in liver microsomal fractions of PB-treated rats and with a reconstituted CYP2B1: NADPH-P450-reductase system are in the range of 3.2D5.0 μм and 2.8D3.5 μм, respectively. Lineweaver-Burk plots with the inhibitors in concentrations that were found to cause roughly a 50% inhibition and with 7-ethoxycoumarin as substrate revealed for all four alkylphospholipids a competitive inhibition type. The degree of the competitive inhibition is quantified by the K i values. With liver microsomal fractions of PB-treated rats, the K i values of rac-2-OH (K i = 1.36 μм) and rac-2-O-acetyl (K i = 1.33 μm) differs slightly from those of rac-2-NH2 (K i = 2.2 μм) and rac-2-N-acetyl (K i = 2.2 μм), but with the reconstituted CYP2B1: NADPHP450- reductase system all K i values are in the small range of 1.8 D 2.6 μм, indicating that the short substituted group at the 2-position (OH; O-acetyl; NH 2 ; N-acetyl) of the long chain octadecanol part of the phosphodiesters exhibit no essential role on the strong inhibitory potency of these alkylphosphocholines on the 7-ethoxycoumarin-O-deethylase activity.

A root culture of Inula helenium L. was established from leaf explants of aseptic seedlings. An ethanol extract from the lyophilised roots was fractionated using different chromatographic techniques (CC, TLC). The main secondary metabolites found in the root culture were two thymol derivatives: 10-isobutyryloxy-8,9- epoxy-thymol isobutyrate (1) and 10-isobutyryloxy-6- methoxy-8,9-epoxy-thymol isobutyrate (2). The compounds were identified by spectral methods. Quantification of compound 1 in plant material was done by analytical RP-HPLC.

trans-Bis(resorcylaldoximato)copper(II) and trans-bis- (2,3,4-trihydroxybenzaldoximato)copper(II) (CuRES 2 and CuTRI 2 , respectively) have been tested for antiviral properties against HIV, using an in vitro assay that measures the ability of the test compounds to prevent the killing of susceptible human cells by HIV. In the case of CuTRI 2 , T4 lymphocytes (CEM-V and CEM-Z cell lines) were exposed to HIV at a virus to cell ratio approx. 0.05 in microtiter plates. In the case of CuRES 2 , a human leukemia cell line (MT-2) was used instead. The tetrazolium salt XTT was added to all wells, and the cultures were incubated and analyzed spectrophotometrically to quantitate formazan production and viewed microscopically for detection of viable cells. In spite of their antiproliferative properties, neither agent had any detectable ability to prevent the cytopathic effects of HIV in cultures of the target cells used. Because the test system employed was constructed in such a way as to detect antiviral agents acting at any stage of the virus reproductive cycle, the results obtained strongly suggest that neither studied agent has any value as the direct prevention of the cell destruction caused by HIV is concerned.