Band 58, Heft 5-6

From dichloromethane extracts of flowerheads of Achillea pannonica Scheele three sesquiterpenes were isolated and identified: 11,13-dehydrodesacetylmatricarin, (6E)-5-tigloxy- 9-hydroxynerolidol and α-longipin-2-en-1-one. The structures were determined by MS, IR and NMR spectroscopic analyses. (6E)-5-Tigloxy-9-hydroxynerolidol is reported here for the first time. Additionally spathulenol, a compound of the essential oil was identified using GC-MS and GC-FTIR.

Cell suspension cultures from hypocotyl-derived callus of Hyssopus officinalis were found to produce two sterols i. e. β-sitosterol (1) and stigmasterol (2), as well as several known pentacyclic triterpenes with an oleanene and ursene skeleton. The triterpenes were identified as oleanolic acid (3), ursolic acid (4), 2α, 3β-dihydroxyolean-12-en-28-oic acid (5), 2α, 3β- dihydroxyurs-12-en-28-oic acid (6), 2α, 3β, 24-trihydroxyolean-12-en-28-oic acid (7), and 2α,3β, 24-trihydroxyurs-12-en-28-oic acid (8). Compounds 5-8 were isolated as their acetates (6, 8) or bromolactone acetates(5, 7)

The essential oils from two Chinese endemic Meconopsis species, i. e., M. punicea and M. delavayi, were analyzed by using GC-MS for the first time. The major constituents were hexadecanoic acid (16.8%), 1,2-dimethyl naphthalene (11.4%), 1,4-dimethyl naphthalene (6.6%), 1,3-dimethyl-5-ethyl naphthalene (5.9%), and 3-methyl biphenyl (5.6%) for M. punicea, and hexadecanoic acid (9.9%), 1,2-dimethyl naphthalene (7.9%), 1,3-dimethyl-5-ethyl naphthalene (6.2%), tetradecane (5.9%), and hexyl cinnamaldehyde (5.5%) for M. delavayi.

The hydro-distilled essential oil from dried aerial parts of one of widespread Iranian skullcaps, Scutellaria pinnatifida A. Hamilt. ssp. alpina (Bornm.) Rech. grown in Khorassan province was analyzed by GC and GC/MS. Thirty components were characterized representing 93.8% of the total components detected. The major components of the oil were germacrene- D (39.7%) and beta-caryophyllene (15.0%).

A Pestalotiopis sp. was isolated from the trunk bark of Pinus taeda. The fungus was cultivated in liquid medium and produced three highly oxygenated caryophyllene sequiterpene derivatives, named pestalotiopsolide A, taedolidol and 6-epitaedolidol, respectively. The sesquiterpenes were isolated by silica gel based chromatographic procedures and their structures were elucidated by NMR spectroscopic data.

The composition of sterols, volatiles and some polar compounds from three Corallina samples (C. granifera and C. mediterranea from the Black Sea and C. mediterranea from the Mediterranean Sea) was established. The sterol composition of the Black Sea samples was similar but it differs from that of the Mediterranean sample. The composition of the volatiles was very complex. The main groups of constituent were hydrocarbons, alcohols, carbonyl compounds, acids and their esters, terpenes. The composition of the polar components, soluble in n-butanol, was also established. There were some differences in the chemical composition of the two Black Sea species, which may be due to the biodiversity between them, while the differences in the composition of the two C. mediterranea samples could be due to the differences in the environment (salinity, temperature, pollution, etc.).

An investigation of the natural products chemistry of the brown alga Sargassum asperifolium from the red sea yielded a new steroidal metabolite, 24-vinyl cholest -4-ene -24- ol -3 one, saringosterone (3), a known steroidal metabolite, 24-vinyl cholest -5-ene -3b, 24-diol, saringosterol (4), and known diterpene with a hydroazulene skeleton, dictyone (1). The identification of the isolated metabolites was established mainly by spectral methods and chemical transformation of the dictyone (1) to its acetate (2).

From the methanolic extract of the underground parts of Globularia cordifolia, a new iridoid glycoside, 5-hydroxydavisioside (1) and a new bisiridoid glycoside, globuloside C (2) were isolated along with six known iridoid glycosides, aucubin, melampyroside, monomelittoside, globularifolin, alpinoside and asperuloside. The structures of the isolates were established by 1D and 2D NMR spectroscopy in combination with IR, UV and MS analyses.

A polysaccharide, an α-ᴅ -glucan with an apparent molecular weight of 6.85×10 4 , called PSa glucan, was isolated from fresh seeds of Sorghum arundinaceum by fractionation on Sephacryl S-300 HR and Sephadex G-25. Chemical and spectroscopic studies indicated that it has a highly branched glucan type structure composed of α-(154) linked d-glucopyranose residues with (1→3), (1→6) branching points, and a significant amount of α-(1→6) branching to α-(1→3) linked ᴅ -glucopyranose residues. The anti-inflammatory activity of the polysaccharide was performed using the capillary permeability assay.

A new highly oxygenated flavone, namely 8, 3′-dihydroxy-5, 6, 7, 4′-tetramethoxyflavone (1), together with other five known flavonoids were isolated from the tropical plant Vernonia saligna. Their structures were established on the basis of spectral (MS, IR, UV, 1D & 2D NMR) measurement and chemical evidence.

A modified in vitro lipid peroxidation inhibition assay was used to guide the fractionation and the isolation of antioxidative principles of Finnish pine bark extract. This approach yielded 3,4-dihydroxybenzoic acid (protocatechuic acid) and taxifolin-3-O-β-glucopyranoside as major antioxidative compounds from the plant material. The structural elucidation of these compounds was undertaken with the help of HPLC-DAD and HPLC-ESI-MS analyses. Their IC 50 values, in comparison to trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), were: trolox (1.78 ± 0.56 μm) < protocatechuic acid (5.77 ± 1.63 μm) < taxifolin-3- O-β-glucopyranoside (16.30 ± 1.98 μm). The method for the determination of antioxidant activity proved reproducible and quick for routine analyses with 96 well plates.

The composition of hexane and ether extracts from buds of two poplar species (Populus balsamifera and P. nigra) was investigated by GC-MS method. In hexane extracts, 54 “neutral” compounds were recorded. The greatest amounts of them are sesquiterpenes and nalkanes. Among 56 components of ether extracts, many aliphatic acids and hydroxyacids were detected. However, the main fraction consists of phenolcarboxylic acids, substituted cinnamic acids, and their esters. It was established that chemotaxonomic differences between Populus balsamifera and P. nigra are observed in the case of both hexane and ether bud extracts.

The antioxidant properties of galantamine hydrobromide ((4α,6β)-4a,5,9,10,11,12-hexahydro- 3-methoxy-11-methyl-6H-benzofuro[3a,3,2-ef][2]benzazepin-6-ol hydrobromide) were studied in vitro, using luminol-dependent chemiluminescence and spectrophotometry. It was found that this compound was a scavenger of reactive oxygen species (ROS). By comparing the antioxidant effects of galantamine ((4α,6β)-4a,5,9,10,11,12-hexahydro-3-methoxy-11-methyl- 6H-benzofuro[3a,3,2-ef][2]benzazepin-6-ol), galantamine hydrobromide, narwedine (4a, 5,9,10,11,12-hexahydro-3-methoxy-11-methyl-6H-benzofuro[3a,3,2-ef][2]benzazepin-6-one), and narwedine hydrobromide it was found that the antioxidant activity depended on the enolic OH group in the molecule. The presence of a quaternary nitrogen in the compound increased the strength of the scavenging effect. It is proposed that the antioxidant properties observed in vitro may contribute to the therapeutical effect of galantamine hydrobromide on patients with brain degeneration.

Synthesis of N- and O-acyl derivatives of ᴅʟ-serine and threo-ᴅʟ-phenylserine was accomplished by a regioselective acylation of the corresponding amino acid. The residues introduced into amino acid structure contain hydrophobic long chain or aromatic, namely lauroyl, myristoyl and phenylacetyl moieties. The fungicidal activity against six strains of fungi was studied. Several compounds were found to be effective against growth of fungi, and Omyristoyl- ᴅʟ-serine 2 and N-phenylacetyl-threo-ᴅʟ-phenylserine 8 completely inhibited the growth of the mycelium of the fungus Verticillium dahliae.

Physiological conditions which lead to changes in total carotenoid content in tomato plantlets were identified. Carotenoid levels were found to increase after the onset of a dark period during a normal 24h cycle. This rapid initial increase is followed by a steady decrease in carotenoid content throughout the night. A decrease in the expression of several carotenogenic genes, namely pds, zds (carotenoid desaturases) and ptox (plastid terminal oxidase), was observed following the removal of the light (when carotenoid content is at its highest). An increase in gene expression was observed before the return to light for pds and zds (when carotenoid levels were at their lowest), or following the return to light for ptox. The phytoene desaturation inhibitor norflurazon leads to a decrease coloured carotenoid content and, in the light, this correlated with pds and zds gene induction. In the dark, norflurazon treatment led to only a weak decrease in carotenoid content and only a small increase in pds and zds gene expression. The striking absence of phytoene accumulation under norflurazon treatment in the dark suggests a down-regulation of carotenoid formation in darkness. However, prolonged dark conditions, or treatment with photosynthetic inhibitors, surprisingly led to higher carotenoid levels, which correlated with decreased expression of most examined genes. In addition to light, which acts in a complex way on carotenoid accumulation and gene expression, our results are best explained by a regulatory effect of carotenoid levels on the expression of several biosynthetic genes. In addition, monitoring of protein amounts for phytoene desaturase and plastid terminal oxidase (which sometimes do not correlate with gene expression) indicate an even more complex regulatory pattern.

The studied problem is of commercial interest because whey, the cultivation substrate, is a waste by-product from the transformation of milk into cheese and casein. Investigations on the influence of the dilution rate (D) on the bioproductivity of lactose-utilizing yeasts were carried out with two model strains D the oxidative strain Candida blankii 35 and the fermentative strain Candida pseudotropicalis 11. The increase of D led to the different changes in productivity. The best synthesizing ability of both continuously cultivated strains is established at D = 0.4 [h-1] despite the different type of metabolism. The oxidative strain C. blankii 35 is more effective in comparison with the fermentative strain C. pseudotropicalis 11 because of its ability to synthesize 1.5 fold higher biomass and protein yields. These experimental facts were proved also by simulative research with a Fuzzy Knowledge-Based System (FKBS) developed for modeling the influence of D on several process variables.

Three types of liposomes in aqueous solution were subjected to a low frequency switched weak magnetic field. A differential non-linear dielectric spectroscopy (DNLDS) was performed at 40 °C with two planar orthogonal electrodes, positioned parallel and vertical to the earth surface. The difference of the free voltage release (FVR) signals for the two orthogonal directions following electric pulses with an amplitude of 1.0 V and a duration of 25 ms, were Fourier-transformed. An additional magnetic field was switched with a period of 400 ms and a variable amplitude from 0 to 100 G, whose direction was parallel to the vertical electrode plane. With two of the liposomes (egg yolk lecithin (EY), asolectin doped with cholesterol (ASCO)) a decrease of the signal amplitude with increasing magnetic fields could be seen in most of the 25 observed harmonic frequencies (relative to the electric pulse frequency f(0) = 40 Hz). For EY liposomes this decrease was highly significant and not linear for the 1.-5., and above the 20. harmonic frequency, ASCO liposomes showed a similar effect. Asolectin liposomes showed the reverse response. Quantum mechanical conditions of charges on the liposome surface are discussed as a possible origin of these effects.

A series of sage phenolics was tested for activity against a panel of Leishmania parasites and for immunomodulatory effects on macrophage functions including release of tumour necrosis factor (TNF), interleukin-6 (IL-6), and interferon (IFN)-like activities. For this, functional bioassays were employed including an in vitro model for leishmaniasis in which macrophage- like RAW 264.7 cells were infected with Leishmania parasites, an extracellular Leishmania growth-inhibition assay, a fibroblast-lysis assay for TNF-activity, a cell proliferation assay using IL-6 sensitive murine B9 hybridoma cells, and a virus protection assay for IFN-like activity. Whereas none of the test samples exhibited marked activities against extracellular Leishmania promastigotes (IC 50 > 700 to > 2800 nм; > 500 μg/ml), caffeic acid, salvianolic acids K and L as well as the methyl ester of salvianolic acid I showed pronounced antileishmanial activities against intracellular amastigote stages within RAW cells (IC 50 3-23 nм vs. 10-11 nм for the reference Pentostam ® ). Noteworthy, the phenolic samples showed no cytotoxicity against the host cells (IC50 > 600 to > 2200 nм; > 400 μg/ml). Tested sage phenolics activated Leishmania-infected RAW 264.7 for release of TNF ranging 22-117 U/ml and IL-6 ranging 3-42 U/ml. In contrast, their TNF- or IL-6-inducing potential in experiments with non-infected host cells was negligible. Furthermore, caffeic acid and salvianolic acid K induced a modest release of IFN-like activity (5-9 and 2-4 U/ml, respectively) as reflected by inhibition of the cytopathic effect of encephalomyocarditis virus on L929 cells. The results support the emerging picture that plant polyphenols may be credited for the profound healthbeneficial properties of various herbal medicines and agricultural products.

The bacterium Propionibacterium acnes is light-sensitive due to porphyrin-induced photosensitization. The light sensitivity increases with incubation of 5-aminolevulinic acid, ALA. For the first time, 1H magic angle spinning NMR spectroscopy is used to describe the photoinduced changes in the bacterium after ALA incubation. Successful photosensitization was performed with light-emitting diodes in the blue and red regions (430 and 654 nm, respectively). The irradiation setup, suitable for irradiation of bacaterium suspensions in petri dishes is described. For NMR studies blue light diodes with about 90 μmol/m2s were chosen. After blue light irradiation, the endogenous glycine betaine, proline, glutamate and choline levels in P. acnes decreased with increasing irradiation time. For sublethal light doses (50% survival fraction), the endogenous glycine betaine level decreased 80% on average. The corresponding percentages for proline, choline and glutamate were about 40, 25 and 10, respectively. It is hypothesized that the irradiation, inducing porphyrin photosensitization amplified by ALA incubation, leads to elimination of the osmolyte glycine betaine and possibly also proline by so-called regulatory volume decrease (RVD) mechanisms. These mechanisms are known to be active in several prokaryotic and eukaryotic cells when exposed to hypotonic stress. They are also known to be present in several eukaryotic cells during photodynamic therapy (PDT) exposure leading to hypotonoc stress. The findings contribute to the knowledge of the inactivation mechanisms of P. acnes in photosensitization, and could therefore be of interest in the efforts to use PDT as treatment of the acne disease.

The sulfurous acid ester, trans-sulfurous acid allyl ester 3-allylsulfanyl-allyl ester 8, along with two known thiosulfinates was isolated from the aqueous ethanol extract of garlic (Allium sativum). The chemical structure of 8 was determined on the basis of spectroscopic data including high resolution mass and two-dimensional NMR techniques. All of these compounds induced methemoglobin formation in a canine erythrocyte suspension in vitro resulting in the oxidation of canine erythrocytes. This is the first report of sulfurous acid ester showing oxidant activity in canine erythrocytes.

We have developed a genetic barcode module, based on a parallel sorting facility of single nucleotide polymorphism for secure individual identification of cattle. Biotinylated allelespecific oligonucleotides were immobilized onto the predefined spots of streptavidin tethered self-assembled monolayers with long chain alkanethiols on biochips. The target DNAs for hybridization and subsequent on-chip minisequencing were produced by multiplex PCR method. After enzymatic extension, only the moiety-modified dideoxynucleotide triphosphate, when coupled to its complementary target sequence, could be detected by the corresponding antibody to the moiety in a specific and sensitive manner. The database SNPZoo was developed for storage of the sequence information consisting of cytosine/thymidine patterns. This SNP chip system can further be used in the detection of any replaceable point mutations occurring in the human and animal genes.

The yellow tortrix, Acleris fimbriana Merick (Lepidoptera: Tortricidae), is an economically important insect pest on fruit trees with four generations a year in North China. In order to develop a new and effective method for forecasting and controlling the pest, the sex pheromone was studied. We have identified the female sex pheromone as (E)-11,13-tetradecadienal (E11,13-14:Ald), (E)-11,13-tetradecadienyl acetate (E11,13-14:Ac) and (E)-11-tetradecenyl acetate (E11-14:Ac). Trapping effect of synthetic chemicals E11,13-14:Ald, alone and in combination with E11,13-14:Ac or/and E11-14:Ac to A. fimbriana males was tested in Beijing Xishan Orchard (2001). E11,13-14:Ald on its own was much attractive to A. fimbriana males. Neither E11,13-14:Ac nor E11-14:Ac alone caught any moths. The catches markedly increased by adding E11,13-14:Ac to E11,13-14:Ald. The optimum ratio of E11,13- 14:Ald and E11,13-14:Ac was 6:4 to 5:5. This attractiveness was apparently enhanced when 5% to 10% of E11-14:Ac was added. The best field activity was in the lure baited with a 6:4:1 ratio of E11,13-14:Ald, E11,13-14:Ac and E11-14:Ac at a dosage of 1000 μg/septum. The effect of antioxidant, 2,6-di-tert-butyl-4-methylphenol [butylated hydroxyluene (BHT)], to the synthetic pheromone blends on its duration and catching efficacy was also tested. Addition of 5-10% BHT to the synthetic pheromone could prolong the life-span of pheromone chemicals for 6-8 weeks, thereby increased its catching efficacy.

Specimens of the spongivorous Mediterranean opisthobranch Tylodina perversa that had been collected while feeding on Aplysina aerophoba were shown to sequester the brominated isoxazoline alkaloids of their prey. Alkaloids were stored in the hepatopancreas, mantle tissues, and egg masses in an organ-specific manner. Surprisingly, the known sponge alkaloid aerothionin which is found only in A. cavernicola but not in A. aerophoba was also among the metabolites identified in wild caught specimens of T. perversa as well as in opisthobranchs with a documented feeding history on A. aerophoba. Mollusc derived aerothionin is postulated to be derived from a previous feeding encounter with A. cavernicola as T. perversa was found to freely feed on both Aplysina sponges in aquarium bioassays. The possible ecological significance of alkaloid sequestration by T. perversa is still unknown.

Eight glucosylceramides (1-8) were isolated from the water-insoluble lipid fraction of a methylene chloride/methanol/water extract of the Patagonian starfish Anasterias minuta. One of the constituents was identified as a new glucosylceramide, anasterocerebroside A (1), while the known glucosylceramide 7 was isolated and characterized for the first time as a pure compound. The structures of 1 and 7 were established by spectroscopic and chemical methods.

β-Sitosteryl-D-glucoside and oleuropein isolated from the olive tree (Olea europaea) and their hydrolysed derivatives were tested by a feeding stimulative activity bioassay using the olive weevil (Dyscerus perforatus). Although the steroidal glucoside showed potent feeding stimulative activity, the activity of the aglycone (β-sitosterol) was significantly lower than that of the glucoside. On the other hand, the difference in the activity between oleuropein, a secoiridoid glucoside, and the hydrolysed derivatives was not significant.

Starvation induced changes in citrate synthase (CS), glucose-6-phosphate dehydrogenase (G6-PDH), lactate dehydrogenase (LDH), DNA, RNA, RNA/DNA ratio and protein were studied in the freshwater catfish Clarias batrachus. Starvation gradually decreased the activity of CS, G6-PDH and LDH in brain, liver and skeletal muscle of the freshwater catfish. The maximum reduction in these enzyme activities upto 35-45% was observed after 35 days of fasting. This shows substantial decline in aerobic and biosynthetic capacity during starvation period. DNA, RNA, RNA/DNA ratio and protein contents were also reduced from 40-67% which reflects reduction in an overall capacity of the protein synthesis. Starvation - induced macromolecular changes indicate impairment of metabolism in fish.

A new p-terphenyl, named aurantiotinin A (1), together with ganbajunin C (2) and atromentin (3) were isolated from the fruiting bodies of the basidiomycete Thelephora aurantiotincta Corner. Their structures were established by spectral (MS, IR, UV, NMR, H-H COSY, HMQC, HMBC measurement) and chemical methods.