Band 63, Heft 11-12

The chemical variations of the essential oil from the aerial parts of Laurus nobilis L. (Lauraceae) have been studied. Plant material has been harvested at each phenological status (vegetative, before anthesis, full flowering and seed-bearing). The oils were obtained by hydrodistillation of the air-dried samples. Analysis by GC and GC-MS of the essential oils has allowed to identify 39 compounds. The main compounds were 1,8-cineole, trans-sabinene hydrate, α-terpinyl acetate, methyl eugenol, sabinene, eugenol and α-pinene.

Ten strains of endophytic fungi were isolated from the bulbs of the traditional Chinese medicinal plant Fritillaria ussuriensis. The extract from one of them, Fu7, showed a positive reaction with Dragendorff’s reagent and the same Rf value in thin-layer chromatography (TLC) analysis as authentic sipeimine. A further TLC scan and high-performance liquid chromatography-evaporative light-scattering detection (HPLC-ELSD) showed that one ingredient of the extract of strain Fu7 had a similar absorption curve in the range 200D700 nm and the same retention time as authentic sipeimine. Thus, the fungus produces the bioactive ingredient sipeimine, as does its host plant, and could be used for the production of sipeimine by fermentation.

The methanolic extract from the barks of the medicinal plant Qualea parviflora (Vochysiaceae) was fractionated by column chromatography over silica gel followed by gel permeation over Sephadex LH-20 to give 3,3′-di-O-methylellagic acid-4-O-β-d-glucopyranoside (1), 3-Omethylellagic acid-4′-O-α-l-rhamnopyranoside (2), 3,3′,4-tri-O-methylellagic acid-4′-O-β-dglucopyranoside (3), and 3,3′-di-O-methylellagic acid (4), together with triterpenes and saponins. We also performed comparative analyses among this species and Q. grandiflora and Q. multiflora using high-pressure liquid chromatography. The biological assays showed that, when compared to the standard ellagic acid, compounds 1-4 are less cytotoxic but have a lower capacity of stimulating murine peritoneal macrophages to release nitric oxide and tumoural-α necrose factor.

Bioguided fractionation of a leaves extract from Limoniastrum feei (Girard) Batt. (Plumbaginaceae) led to the isolation of seven polyphenolic constituents: gallic acid (1), myrciaphenone A (2), myricetin-3-O-β-galactopyranoside (3-1), epigallocatechin gallate (3-2), myricetin 3-O-α-rhamnopyranoside (4), quercetin (5) and myricetin (6). Gallic acid was the most antioxidant compound in DPPH [(0.94 d 0.68) μg/mL] and FRAP [(0.83 d 0.15) μm Fe 2+ /mL] tests, whereas myricetin was a more specific superoxide radical scavenger since it was the most active product in the superoxide nitroblue tetrazolium hypoxanthine/xanthine oxidase test [(1.86 d 0.12) μg/mL].

Rotundifolone, a monoterpene isolated from the essential oil of the leaves of Mentha x villosa, is a constituent of several essential oils and known to have spasmolytic activity. The present study aimed to investigate the correlation between structure and spasmolytic activity of rotundifolone and its analogues in ileum isolated from guinea-pig. Five of the seven tested analogues were found to have a spasmolytic effect more potent than rotundifolone itself, except for pulegone and (+)-limonene. The comparison between rotundifolone and limonene oxide showed that the absence of the keto group did not decrease the relaxant effect. Comparison of the spasmolytic activity between rotundifolone and (+)-pulegone showed that the absence of the epoxy group did not decrease the relaxation of the ileum. Carvone epoxide was found to be significantly more potent than rotundifolone. The monoterpene (-)-carvone produced ileum relaxation and was more potent than its enantiomer (+)-carvone. (+)-Limonene and pulegone oxide showed a similar effect. The study showed that the functional groups and their position at the ring of rotundifolone contributed to the relaxation activity of the ileum. The absence of the oxygenated molecular structure is not a critical requirement for the molecule to be bioactive.

Together with two known cycloartane-type glycosides, askendosides D (3-O-[α-arabinopyranosyl-( 1→2)-β-xylopyranosyl]-6-O-β-xylopyranosyl-cycloastragenol, 2) and G (3-O- [α-arabinopyranosyl-(1→2)-β-xylopyranosyl]-16-O-β-glucopyranosyl-3β,6α,16β,24(R),25- pentahydroxycycloartane, 3), also a new monodesmosidic cycloartane-type glycoside, elongatoside (1), was isolated from the roots of Astragalus elongatus and identified as 3-O- [α-arabinopyranosyl-(1→2)-β-xylopyranosyl]-cycloastragenol. All structures were unambiguously determined by means of spectroscopic and spectrometric methods (1D and 2D NMR, ESI-MS). The isolated compounds were tested for the inhibition of proliferation and ICAM-1 expression in vitro using the human microvascular endothelial cell line (HMEC-1). 1 showed weak activity in the ICAM-1 assay.

A chloroform extract from roots of Craniolaria annua provided six new C-11 unsubstituted abietanes, 14-hydroxy-6,12-dione-7,9(11),13-abietatriene (1), 14-hydroxy-12-oxo-7,9(11),13- abietatriene (2), 6,12,14-trihydroxyabieta-5,8,11,13-tetraen-7-one (3), ar-abietatriene-12-ol- 6,7-dione-14,16-oxide (4), ar-abietatriene-12,16-diol-14,16-oxide (5) and ar-abietatriene-12-ol- 7-one-14,16-oxide (6), and two known compounds, ferruginol and stigmasterol. The structures of both the new and known compounds were established by spectroscopic methods. Abietane 1 gave 14-hydroxy-6-oxoferruginol (1A) upon treatment with NaBH 4 . Abietanes 1, 1A, 3-5 and ferruginol showed cytotoxic effects against trypomastigote and epimastigote forms of Trypanosoma cruzi and against fibroblastic Vero cells.

Chalcones belong to a class of α,β unsaturated aromatic ketones which occur abundantly in nature, especially in plants. They are promising and interesting compounds due to their vast applications in pharmaceuticals, agriculture and industry. Several studies have shown that these compounds exert important biological activities in different experimental models. The present work deals with the antinociceptive activity, evaluated against the writhing test, of three series of chalcone-like compounds obtained by the Claisen-Schmidt condensation, using different aldehydes and substituted acetophenones. The results reveal that the compounds synthesized show a significant antinociceptive effect compared with nonsteroidal drugs such as aspirin, paracetamol and diclofenac. They also show that the electronic demand of the substituents is the dominant factor of the biological activity.

The furanoeremophilanes 6β-angeloyloxy-1, 10-dehydrofuranoeremophilan-9-one (1), 6β- hydroxy-1, 10-dehydrofuranoeremophilan-9-one (2) and 6β-propionyloxy-1, 10-dehydrofuranoeremophilan- 9-one (3) were isolated from Senecio otites, their structures elucidated by spectral analyses, and their insecticidal and phytotoxic properties evaluated. Compounds 1-3 proved to be effective aphid antifeedants against Myzus persicae and Rhopalosiphum padi and had postingestive negative effects on Spodoptera littoralis larvae. These compounds did not have any phytotoxic effects on Lactuca sativa.

1-Phenylhepta-1,3,5-triyne (1), 5-phenyl-2-(1′-propynyl)-thiophene (2), and 2-(3′-acetoxy- 1′-propynyl)-5-phenylthiophene (3) were isolated from Coreopsis lanceolata L., and their structures identified by spectroscopic methods. Compounds 1 and 2 showed effective nematicidal activities against Bursaphelenchus xylophilus and Caenorhabditis elegans, but had hardly any effect against Pratylenchus penetrans. Compound 3 did not show any effective nematicidal activity.

In the course of our search for antimalarial leads from marine algae, four metabolites, sargaquinoic acid, sargahydroquinoic acid, sargaquinal and fucoxanthin, were isolated from the South African alga Sargassum heterophyllum. Fucoxanthin and sargaquinal showed good antiplasmodial activity toward a chloroquine-sensitive strain (D10) of Plasmodium falciparum (IC 50 1.5 and 2.0 μm, respectively), while sargaquinoic acid and sargahydroquinoic acid were only moderately active (IC 50 12.0 and 15.2 μm, respectively).

A series of agarofuran compounds isolated from Chilean Celastraceae were evaluated in vitro for their cholinesterase inhibitory activities by UV spectroscopy.

Mutations in the p53 tumour suppressor gene have been associated with chemical carcinogens. Natural antimutagens are promising modulators for reducing the cancer risk. The present study was carried out to assess the protective efficacy of some natural antimutagens against p53 alterations. We investigated the ability of curcumin (100 mg/kg BW) and chlorophyllin (3 mg/kg BW) pretreatment, for three times per week for three successive weeks, to inhibit mutations induced by intraperitoneal injection of a single dose of 40 mg/kg BW of cyclophosphamide (CP). Forty male albino rats were assigned into four groups: control nontreated group, CP-treated group, curcumin-CP-treated group, and chlorophyllin-CP-treated group. Liver samples were collected for DNA isolation two days after CP injection. The isolated DNA was used in single-strand conformational polymorphism (SSCP) analysis of polymerase chain reaction (PCR)-amplified products of four regions: two in exon 5, one in exon 6, and one in exon 7. The amplified products of p53 different regions were found to be in the expected molecular size of the designed primers. SSCP analysis of these amplified products showed that CP-induced mutation in the p53 gene was found only in exon 7 shifting its electrophoretic mobility. Chlorophyllin treatment prior to CP injection had a more potent protective efficacy (80%) than that with curcumin (33.3%).

A field study was conducted during 1994 to 1998 on the Experimental Farm Roggenstein, near Fürstenfeldbruck, Bavaria, Germany to determine the effect of transgenic glufosinateresistant rape in combination with the herbicide Basta ® [glufosinate-ammonium, phosphinothricin, ammonium (2RS)-2-amino-4-(methylphosphinato) butyric acid] application on soil microorganisms and the behaviour of the synthetic transgenic DNA in response to normal agricultural practice. No influence of Basta ® on microbial biomass could be detected. The phospholipid fatty acid analysis of soil extracts showed no difference between Basta ® application and mechanical weed control, whereas conventional herbicide application revealed a different pattern. Basta ® application resulted in a changed population of weeds with a selective effect for Viola arvensis. During senescence, transgenic rape DNA was degraded similar to endogenous control DNA. After ploughing the chopped plant material in the soil, transgenic as well as endogenous control DNA sequences could be detected for up to 4 weeks for rape and up to 7 months for maize, whereas PCR analysis of composted transgenic maize revealed the presence of the transgene over a period of 22 months.

The tolerance, sorption of nickel(II) ions, and changes in the production and composition of exopolymers of eight yeast strains grown under nickel presence with/without NaCl were studied. Strains of Pichia anomala and Candida maltosa known as the most resistant yeasts against nickel tolerated up to 3 mm Ni 2+ . NaCl addition decreased both the resistance of the yeast strains toward nickel ions and the sorption of metal ions into cells. All yeasts absorbed nickel predominantly into exopolymers (glycoproteins) and on the surface of cells. However, while the amount of polysaccharide moieties of exoglycoproteins of most of the resistant yeasts was induced by stress conditions, the ratio polysaccharide/protein in the exopolymers remained unchanged in the sensitive species Cystofilobasidium. The exopolymer composition might play a key role in yeast adaptation to stress conditions caused by heavy metal ions.

Bacopa monnieri (L.) Wettst. was studied for shoot induction and regeneration on Murashige and Skoog (MS) medium supplemented with different plant growth regulators. Stem explants cultured on medium containing 0.1 mg/l thidiazuron (TDZ) resulted in the highest number of shoots (117 shoots/explant). Regenerated plants from medium with 0.5 mg/l TDZ contained the highest level of pseudojujubogenin glycosides [(30.62 d 1.29) mg/g dry wt] which was 2-fold higher than that of in vitro grown plants of the same age [(16.96 d 1.49) mg/g dry wt]. Plantlets regenerated from 0.1 mg/l TDZ also showed a high level of pseudojujubogenin glycosides [(27.94 d 1.19) mg/g dry wt]. The effect of elicitor on pseudojujubogenin glycosides accumulation in B. monnieri whole plant cultures was investigated. Chitosan at 150 mg/l and yeast extract at 2 mg/ml increased the pseudojujubogenin glycosides production [(40.83 d 2.24) mg/g dry wt and (40.05 d 2.37) mg/g dry wt, respectively] after 7 days, which was 6-fold higher than in the control cultures.

CoQ 10 is used not only as a medicine but also as a food supplement due to its various physiological activities. The production of CoQ 10 by microbes is a successful approach for generating large amounts of this natural product. The effects of dissolved oxygen (DO) contents and the two-stage fermentation process on cell growth and CoQ 10 production by Rhodopseudomonas palustris J001 were investigated. The optimal DO contents for cell growth and CoQ 10 production were 45% and 15%, respectively. A two-stage fermentation process, which consists of a 1 st stage with 45% DO, a 2 nd stage with 15% DO and a synchronous feeding of 2.0% NaAc at the switching time (42 h after inoculation), has proven to be the optimum fermentation process for the production of CoQ 10 . The maximum biomass, CoQ 10 production and CoQ 10 production rate were 1.31 g l -1 , 89.1 mg l -1 , and 1.142 mg l -1 h -1 , respectively, increased by 28%, 585% and 426% as compared to the one-stage batch production with 45% DO. The DO level was the major factor to increase the CoQ 10 production by the two-stage process.

Dielectric measurements in the frequency range 10 5 -10 8 Hz were performed on wild-type (wt) adenosylribosyl transferase and a mutant enzyme. The analysis of the dielectric relaxation curve allowed the estimation of the hydrodynamic radius and of the electric dipole moment. The first parameter remained unchanged in wt and mutant protein. The dipole moment of the mutant, however, was significantly increased. Implications on the electrostatic interactions between enzyme and substrate are discussed.

α-Galactosidase production by the fungus Humicola lutea 120-5 immobilized in a hybrid sol-gel matrix, consisting of tetraethylorthosilicate (TEOS) as a precursor and a mixture of polyethyleneglycol (PEG) and polyvinylalcohol (PVA), was investigated under semicontinuous shake flask cultivation and compared to the enzyme secretion by free cells. The influence of the carrier weight on the α-galactosidase biosynthesis in repeated batch experiments was followed. Best results were obtained with 2 g of the sol-gel particles per culture flask using 144-h runs. The growth behaviour of the immobilized mycelium during both the growth and productive phases was observed by scanning electron microscopy. The presence of abundant mycelial growth of intact hyphae correlated with a 2-fold higher enzyme activity compared to free cells. The obtained biocatalyst retained a high level of enzyme titer exceeding the activity of free cells during four cycles of operation (24 days). This result is confirmed by the micrographs showing the retained viability of the growing vegetative cells due to the protective role of the carrier.

The base sequences of the nucleic acids corresponding to ten proteins (aconitase, alcohol dehydrogenase, enolase, fumarase, isocitrate dehydrogenase, lactate dehydrogenase, phosphofructokinase, phosphoglycerate mutase, pyruvate kinase and succinate dehydrogenase) belonging to a total of 154 species, ranging from prokaryotes to vertebrates, were compared with the base sequences of oligoribotides whose growth rates were calculated by a chemical kinetics model. It was shown that oligoribotides grown according to the kinetics model have a fraction of repetitive bases larger than expected from random processes. The base sequences of nucleic acids of prokaryotes and eukaryotes retain, in decreasing proportions, this feature of their abiotic past. Chemically synthesized pentameric stretches with repetitive bases are slightly more abundant than those present in prokaryotes. Genetic drift and natural selection, operating as fundamental laws even for the most primitive living systems, reduced the original, chemically controlled, repetitive base frequency in prokaryotes, which was further reduced for eukaryotes.

We searched and analyzed cyanobacteria-originated metazoa/fungi proteins (COPs) by phylogenetic analyses. Analysis of them showed that for millions of years universal plastid endosymbiosis and gene transfer occurred in ancestors of metazoa/fungi, and some transferred fragments have been reserved until now even in modern mammals. Most eukaryotes contained plastids in the ancient era, and some of them lost them later. Functions of homologues in cyanobacterial genomes and eukaryotic genomes are in consensus, and most are involved in the organic compound metabolism. With emergence of organelles and subcellular structure in eukaryotic cells, the locations of these proteins diversified. Furthermore, some novel functions were endowed for COPs, especially in vertebrates.

Antifungal properties of rabbit antiserum prepared by immunization are reported. The immunization was done by a chemically prepared conjugate consisting of Candida albicans (serotype A) surface mannan and human serum albumin. Addition of rabbit antiserum to d-glucose medium inoculated with C. albicans effectively inhibited its growth. Moreover, C. albicans cells treated with rabbit antiserum revealed the entire loss of viability (expressed as decreased mitochondrial dehydrogenase activity). No growth of treated cells on an agar plate was observed. The results confirmed that the mannan-protein conjugate could be considered as an effective component of perspective vaccine.

Acrylamide is a reactive neurotoxin with a high intestinal bioavailability. Recently we have shown that under the pH regime of the gut acrylamide can react with proteins and that this reaction reduces the uptake of acrylamide in a gut model. On the other hand, using radioactive labeled acrylamide, Bjellaas et al. [Toxicol. Sci. 100, 374-380 (2007)] showed that in vivo the vast majority of orally administered acrylamide is absorbed and excreted as N-acetyl-S- (3-amino-3-oxopropyl)-cysteine with the urine. Therefore, we tested whether intestinal proteases can degrade a protein with acrylamide bound to cysteine residues. Furthermore we tested whether the product of this reaction, S-(3-amino-3-oxopropyl)-cysteine, can pass the intestinal barrier. Here we showed that S-(3-amino-3-oxopropyl)-cysteine is indeed a product of proteolytic degradation of acrylamide-treated proteins. Using Caco-2 cells as a gut model, we further showed that the non-protein amino acid S-(3-amino-3-oxopropyl)-cysteine is a substrate for the neutral and cationic amino acid transporter system. Hence we concluded that protein-bound acrylamide can be released in the intestine and that the resulting product S-(3-amino-3-oxopropyl)-cysteine is transported through the intestinal barrier and later excreted via the urine.

Yeast cell wall matrix particles are composed entirely of β-glucan and mannoprotein. Alkali-insoluble (1→3)-β-d-glucan was extracted from the yeast cell wall by an alkaline-acid method. IR spectra analysis showed that the product was chemically pure glucan, that is to say, it contained no other carbohydrates and proteins. So, the alkaline-acid method was ideal for extracting (1→3)-β- d-glucan from the yeast cell wall. We also purified and analyzed mannan oligosaccharides by the dilute alkali- Sevage method from the yeast cell wall.

d-(+)-Pinitol, a natural product of the group of cyclitols, was purified for the first time from an aqueous extract of the heartwood of Enterolobium cyclocarpum, and its chemical structure was determined.