Volume 55, Issue 3-4

Native and peracetylated sporopollenin from the pollen of Typha angustifolia L. was investigated using several spectroscopic methods, inducing Fourier transform infrared spectroscopy (FTIR), solid-state 13 C-nuclear magnetic resonance spectroscopy ( 13 C-NMR) and Xray photoelectron spectrometry (XPS). Interpretation of the experimental data shows that the greater part of oxygen found in sporopollenin originates from hydroxyl groups and must be derived from aliphatics and not from aromatics. This result indicates that not only aromatics and long unbranched aliphatics but also poly-hydroxyl aliphatic components are involved in the complex structure of the polymer. Furthermore, it is most probable that the monomers of the sporopollenin skeleton are linked by ether- and not by ester-linkage. Two possible approaches are suggested for the characterisation of sporopollenin structure.

A new iridoid glucoside, 5-desoxysesamoside, was isolated from Phlomis tuberosa L. (Lamiaceae) together with three known iridoid glucosides sesamoside, shanziside methyl ester and lamalbid. Lamiide was found in P. herba-ventis ssp. pungens in high concentrations.

A novel cucurbitacin glycoside has been isolated from aerial parts of Kageneckia oblonga R. et P. and shown to be 3β-(β-D-glucosyloxy)-16α,23α-epoxycucurbita-5,24-dien-11-one. The structure was established by usual spectroscopic and two-dimensional (2D ) NMR techniques. This compound has found to be nontoxic when tested in-vivo cell culture assays. In previous investigations we reported 23,24-dihydrocucurbitacin F and prunasine. This was the first report on cucurbitacins from the genus Kageneckia (Rosaceae).

Pseudomonas spp. of the fluorescent group produce siderophores (so-called pyoverdins) consisting of a peptide chain attached to a pyrimidoquinoline ring system which is derived from a condensation product of L-Dab and D-Tyr. Commonly several related compounds are found to accompany the pyoverdins having the same peptide chain, but differing in the heterocyclic part. The structure elucidation of a new variety (succinopyoverdin) is described here.

The structure which had been proposed for the pyoverdin named pseudobactin 7SR1 (Yang and Leong, 1984) differed from those of all other pyoverdins investigated sofar: its peptide chain was supposedly linked to the chromophore not by an amide bond originating from its N-terminal amino acid, but rather by an ester bond involving one of the three Ser. It will be shown that the peptide chain of pseudobactin 7SR1 is actually bound to the chromophore amidically by its N-terminal Ser and that it comprises a cyc/odepsipeptidic substructure with an ester bond between the C-terminal Thr and the OH-group of the second Ser in the chain.

Two novel isoflavonoid dimers presumably originating from 2′-hydroxygenistein, 5,7,4′- trihydroxycoumaranochroman-4-one-(3→5‴)-5″,7″,2‴,4‴-tetrahydroxyisoflav- one (1, lupinalbisone A ) and 5,7,4′-trihydroxycoumaranochro-man-4-one-(3→6″)-5″,7″,2‴,4‴-tetrahydroxyisoflavone (2, lupinalbisone B) were isolated from the roots of Lupinus albus L., and their structures involving relative stereochemistry were elucidated by spectroscopic methods. Using horse radish peroxidase and 2′-hydroxygenistein (3) as the substrate revealed the formation of these dimers together with 5,7,4′-trihydroxycoumaronochromone (4, lupinalbin A ). Dimerization of 3 caused a remarkable increase of antifungal activity.

The fractions of hexane and dichloromethane extraction from marupá (Simaruba amara) and (Bertholletia excelsa) leaves were analyzed by HT-HRGC (high temperature high resolution gas chromatography) and HT-HRGC coupled to mass spectrometry (HT-HRGC-MS). Several compounds can be characterized including unusual high molecular weight com pounds.

Commonly used dynamic sorption techniques for collecting biologically active volatile compounds have been compared. Solid phase microextraction (SPME) using two types of fibers (polydimethylsiloxane, PDMS, 100 μm, and carbowax/divinylbenzene, CW/DVB, 65 μm) was compared to purge and trap methods (Porapak Q, Tenax TA and charcoal) and a technique based on absorption in methanol in a cooling bath. Sampling was done in a stream of purified air (20 ml/min) in a closed and temperature-regulated (27 °C) glass tube, passing over a capillary tube containing a hexane solution of tridecane, heptadecane, 1-octen-3-ol, 1- hexadecanol, ethyl tetradecanoate, α-pinene, linalool, terpinen-4-ol, cis-verbenol, verbenone, β-caryophyllene, E,E-farnesol, and geranylgeraniol. With all of the methods, the sampling was performed for a period of 30 min before extraction and analysis was done on a GC-FID system. In general, SPME gave a higher response for all compounds except for a-pinene, which was only extracted by the CW/DVB fiber. Purge and trap methods and methanol absorption gave the same response for all substances extracted. None of the methods extracted hexadecanol and geranylgeraniol under the conditions used. However, the SPME equipped with the PDMS coating extracted heptadecane, E,E-farnesol and ethyl tetradecanoate. Our results show that SPME, when selecting the fibers to fit the polarity and volatility of the compounds, is an outstanding extraction method compared to purge and trap and methanol absorption, especially for a qualitative analysis. The best conditions for storing fibers exposed to compounds of high volatility were at low temperatures (6 °C) in sealed vials, while the worst way was to leave the exposed fiber unprotected at room temperature (22 °C). The dynamic sampling system was effectively tested on a fruiting body of a polypore fungus ( Ganoderma applanatum) emitting 1-octen-3-ol, and again SPME showed to be the most sensitive technique.

The kernels of Trichosanthes kirilowii seeds contain a green oil which makes up for 62% of their dry matter. This oil consists up to 95% of triglycerides, 2% of glycolipids, 1.3% of phospholipids and 1.8% of chlorophylls. As fatty acid components the triglycerides, glycolipids and phospholipids contain the unsaturated fatty acids linoleic and oleic acid and the saturated palmitic acid. In the triglycerides 19% of the C 18:3 acid occur with the configuration Δ 9 cis, Δ 11 trans. Δ 13 cis. This acid is called trichosanic acid and is absent in glycolipids and phospholipids which contain instead another C 18:3 fatty acid, which has conjugated double bounds and occurs with an amount of 21% and 3%, respectively. Typically, these oil seeds contain in addition up to 30% of their dry matter proteins and up to 2.5% mono- and oligosaccharides. The monosaccharides consist of rhamnose, galactose and glucose and the oligosaccharides represent a mixture of tri- and tetrasaccharides.

A new two-dimensional TLC technique was developed to separate substituted anthraquinones on silica plates using n-pentanol-pyridine-methanol (6:4:3, v/v/v) and toluene-ethyl acetate-ethanol-formic acid (10:8:1:2, v/v/v/v) as eluents. The good separation power of the new technique was demonstrated by applying it to the analysis of complex anthraquinone mixtures isolated from the Scandinavian Derm ocybe sanguinea. Emodin, physcion, endocrocin, dermolutein, dermorubin, 5-chlorodermorubin, emodin-1-β-D-glucopyranoside, dermocybin-1-β-D-glucopyranoside and dermocybin, and five new, earlier in D. sanguinea unidentified compounds, 7-chloroemodin, 5,7-dichloroemodin, 5,7-dichloroendocrocin, 4-hydroxyaustrocorticone and austrocorticone, were separated and identified on the basis of R f values, UV/Vis spectra and mass spectra.

Axenic mycelia of the ectomycorrhizal basidiomycete, Suillus bovinus, were grown in liquid media under continuous aeration with compressed air at 25 °C in darkness. Provided with glucose as the only carbohydrate source, they produced similar amounts of dry weight with ammonia, with nitrate or with alanine, 60-80% more with glutamate or glutamine, but about 35% less with urea as the respectively only exogenous nitrogen source. In crude extracts of cells from NH 4 + -cultures, NADH -dependent glutamate dehydrogenase exhibited high aminating (688 nmol x mg protein -1 × min -1 ) and low deaminating (21 nmol x mg protein -1 × min -1 ) activities. Its K m -values for 2-oxoglutarate and for glutamate were 1.43 mᴍ and 23.99 mᴍ, respectively. pH-optimum for amination was about 7.2, that for deamination about 9.3. Glutamine synthetase activity was comparatively low (59 nmol x mg protein - 1 × min -1 ). Its affinity for glutamate was poor (K m = 23.7 mᴍ), while that for the NH 4 + replacing NH 2 OH was high (K m = 0.19 mᴍ). pH-optimum was found at 7.0. Glutamate synthase (= GOGAT) revealed similar low activity (62 nmol x mg protein -1 × min -1 ), K m -v alues for glutamine and for 2-oxoglutarate of 2.82 mᴍ and 0.28 mᴍ, respectively, and pH-optimum around 8.0. Aspartate transaminase (= GOT) exhibited similar affinities for aspartate (K m = 2.55 mᴍ) and for glutam ate (K m = 3.13 mᴍ) , but clearly different K m -values for 2-oxoglutarate (1.46 mᴍ) and for oxaloacetate (0.13 mᴍ) . A ctivity at optim um pH of about 8.0 was 506 nmol x mg protein -1 × min -1 for aspartate conversion, but only 39 nm ol x mg protein -1 × min -1 at optimum pH of about 7.0 for glutam ate conversion. A ctivity (599 nm ol x mg protein -1 × min -1 ), substrate affinities (K m for alanine = 6.30 mᴍ , for 2-oxoglutarate = 0.45 mᴍ) and pH-optimum (6 .5 -7 .5 ) proved alanine transaminase (= GPT) also im portant in distribution o f intracellular nitrogen. There was comparatively low activity of the obviously constitutive enzyme, urease, (42 nm ol x mg protein -1 × min -1 ) whose substrate affinity was rather high (K m = 0.56 mᴍ). Nitrate reductase proved substrate induced; activity could only be measured after exposure of the mycelia to exogenous nitrate. Routes of entry of exogenous nitrogen and tentative significance of the various enzymes in cell metabolism are discussed.

In this study we used the yeast Candida guilliermondii FTI 20037 immobilized by entrapment in Ca-alginate beads (2 .5 -3 mm diameter) for xylitol production from concentrated sugarcane bagasse hemicellulosic hydrolysate in a repeated batch system. The fermentation runs were carried out in 125- and 250-ml Erlenmeyer flasks placed in an orbital shaker at 30 °C and 200 rpm during 72 h, keeping constant the proportion between work volume and flask total volume. According to the results, cell viability was substantially high (98%) in all fermentative cycles. The values of parameters xylitol yield and volumetric productivity increased significantly with the reutilization of the immobilized biocatalysts. The highest values of xylitol final concentration (11.05 g/1), yield factor (0.47 gig) and volumetric productivity (0.22 g/lh) were obtained in 250-ml Erlenmeyer flasks containing 80 ml of medium plus 20 mi of immobilized biocatalysts. The support used in this study (Ca-alginate) presented stability in the experimental conditions used. The results show that the use of immobilized cells is a promising approach for increasing the xylitol production rates.

From the marine sponge Callyspongia aerizusa collected from the Sea of Bali, Indonesia, fungal isolates of Drechslera hawaiiensis were obtained. Culture filtrates of the fungus yielded four spiciferone derivatives which include spiciferone A (1) and B (2), and two other novel derivatives including spiciferol A (3) which is an alcohol congener of spiciferone A (1) and compound 4 which is an monocyclic spiciferone congener featuring a butoxyl side chain. The structures of the novel compounds were established on the basis of NMR spectroscopic ( 1 H , 13 C, COSY) and mass spectrometric (EIMS) data.

Alfalfa plants (Medicago sativa cv. Europe) inoculated with Sinorhizobium meliloti 2011 (formerly Rhizobium meliloti, de Lajudie et al., 1994) were cultivated for 14 days under standardized growth conditions in mineral medium with addition of the heavy metal cadmium or the polycyclic aromatic hydrocarbon fluoranthene. These xenobiotics significantly reduced the numbers of root nodules before any visible damage to the plant could be detected. EC10. EC50, and EC90 (effective concentrations reducing nodulation, shoot and root fresh weight by 10, 50, or 90% compared to the control without pollutant) were calculated. EC50 for cadmium ranged from 5.8 jam (nodulation) to more than 20 μᴍ (root fresh weight). Testing fluoranthene resulted in an EC50 of 2.5 μg cm -2 for nodulation, and EC50 values of more than 35 μg cm -2 for shoot and root biomass production, indicating that the effect parameter nodulation is 10-fold more sensitive than shoot and root fresh weight. With m RNA differential display techniques the effects of both xenobiotics on gene expression in alfalfa root systems were studied. 37 differentially displayed transcripts were detected. Two of them, called DDMs1 and DDMs2, were confirmed by northern hybridization to be down-regulated in the presence of the xenobiotics. The expression of transcript DDMs1 was enhanced in alfalfa control plants inoculated with rhizobia, the transcript level was increased 2.5-3-fold compared to non-inoculated plants. This positive effect of nodulation was suppressed, partly by 35 μg cm -2 fluoranthene and totally by 20 μᴍ cadmium. The decrease in DDMsl transcription was highly affected by the cadmium concentration with an EC50 of 5.9 μᴍ . Compared to nodulation, almost identical EC10, EC50. and EC90 values were found for DDMsl expression. Sequence analysis of DDMsl revealed a significant overall homology (50% identity) to a hypothetical protein from Arabidopsis thaliana with high similarity to a copper transporting ATPase. High levels of transcript DDMs2 were observed in control plants with a 50% decrease in the xenobiotic-treated plants. DDM s2 gave a strong homology (82% identity) to the cytoplasmatic 60S ribosomal protein L18 from Arabidopsis thaliana.

After feeding of 24-epi-castasterone to the cockroach Periplaneta americana an organspecific epimerization of the brassinosteroid to 2,24-depi-castasterone could be detected in female insects. The metabolite being observed only in the ovaries and not in the testes of the insect was identified by GC/MS in comparison with a synthesized authentic sample. Contrary, 24-epi-brassinolide is not metabolized in the sexual organs of Periplaneta americana. This is the first evidence of a metabolic transformation of a brassinosteroid in insects.

Effects of dodecyltrimethylammonium chloride (DTAC), dodecyltrimethylammonium bromide (DTAB) and dodecyltrimethylammonium iodide (DTAI) on thermotropic phase behaviour of phosphatidylcholine bilayers as well as on 1 H NMR spectra were studied. In order to enhance the effect of counterions on water structure two series of experiments were performed. In the first one the surfactants were added to the water phase and in the other one directly to lipid phase (a mixed film was formed). The effects of particular surfactants on the main phase-transition temperature were more pronounced when they were added to the water phase (1 st method) instead of the lipid phase (2 nd method). Furthermore, in the case of the first method the transitions were found asymmetrical while in the second method nearly symmetrical. It is suggested that surfactant-poor and surfactant-rich domains are formed when surfactants are added to the water phase.

Polar lipids of the cephalopods Eledone moschata, Sepia officinalis and Todarodes sagittatus mantle, represent 50.5%, 66.1% and 74.2% of wet tissue respectively. On the other hand the polar lipids of these three species of cephalopods constitute of 80.8%, 94.8% and 93.7% of phospholipids, respectively. The main phospholipids identified were phosphatidylcholine (52.2, 51.3 and 58.4% of total phospholipids respectively in the above mentioned species), phosphatidylethanolamine (18.1, 19.7 and 23.9%), sphingomyelin (10.7, 15.2 and 6.7%), lysophosphatidylcholine (3.1, 3.8 and 1.8%) and the unusual lipid ceramide aminoethylphosphonic acid (15.9, 10 and 9.2%). The 56.8% of phosphatidylcholine in Eledone moschata, the 46% in Sepia officinalis and the 74.1% in Todarodes sagittatus refer to the structure of 1,2-diacyl-glycerocholine and the remaining percentage refer to the structure of 1-o-alkyl-2-acyl-glycerocholine or 1-o-alkyl-lenyl- 2-acyl-glycerocholine. The 87.2% of phosphatidylethanolamine in Eledone moschata, the 81% in Sepia officinalis and the 90.7% in Todarodes sagittatus refer to the structure of 1,2-diacyl-glyceroethanolamine and the remaining percentage refer to the structure of 1-o-alkyl-2-acyl-glyceroethanolamine or 1-o-alkyl-1-enyl-2-acyl-glyceroethanolamine. The major saturated fatty acids in phosphatidylcholine and phosphatidylethanolamine were C16:0 (30.3-67.5% and 23.2-54.5% ) and C18:0 (3.6-17% and 15.4-28% ), respectively, while the major unsaturated fatty acids in these lipids were C18:1n-9, n-7 (1.0-7.3% and 5.3-10.5% ), C20:5n-3 (1.5-9.8% and 4,5-15.8% ) and C22:6n-3 (12.5-42.0% and 7 .0 - 11.3%), respectively.

The antinociceptive effects of morusin (1), the main prenylflavonoid present in the Morns nigra root barks have been investigated in classical models of pain in mice. The results showed that 1 exhibits a prom ising antinociceptive or analgesic profile by the intraperitoneal route, being more potent than some standard drugs used as reference. The mechanism by which the morusin exerts antinociceptive activity still remains undetermined, but our results strongly suggest that it involves the participation of the opioid system.

Galectin-3 is a β-galactoside-binding lectin that has been implicated in numerous physiological processes, including mRNA splicing, cell differentiation, tumor metastasis and the stress response. We have studied effects of transfer of resident murine peritoneal macrophages to in vitro conditions on galectin-3 in different cell compartments. Galectin-3 was purified by immunoprecipitation with rat monoclonal antibody M3/38, and analyzed by immunoblotting using the same antibody. Transfer to in vitro conditions nearly doubled the total amount of galectin-3 in cells, and caused significant alterations in its intracellular distribution, indicating that galectin-3 is involved in the adaptation of peritoneal macrophages to in vitro conditions.

In previous studies it has been shown that exposure of mice to a 12-Hz 6 mT unipolar square pulsed magnetic field (PMF) suppressed the excess of weight due to application of 1st cold-pressure sunflower oil. This time we considered the effect of oil and/or PMF on the growing curves lifespans of mice. The exposure took place for 30 min 5 days a week, from the 7th week of life to death. The results are 1) a broken slope in the growing curves from the 125th day of aging: the exposed mice were lighter than the controls, keeping the differences between the growing curves needed a repeated exposure all life long; 2) a significant increase in the lifespan of the controls which received oil versus the controls which received water; 3) an increase in the lifespan of the exposed mice versus the non-exposed control batches. On one hand it has been reported that essential polyunsaturated fatty acids found in first cold-pressure sunflower oil played a prominent role in membrane structures and in immune equilibrium. On the other hand, it was shown that oscillating electric fields could activate Na + ,K + -ATPase.

The aim of this study was to investigate whether the preincubation of brain homogenates with L-phenylalanine (Phe), L-cysteine (Cys) or reduced glutathione (G SH) could reverse the free radical effects on Na + ,K + -ATPase activity. Two well established systems were used for the production of free radicals: 1) FeSO 4 (84 μᴍ ) plus ascorbic acid (400 μᴍ ) and 2) FeSO 4 , ascorbic acid and H 2 O 2 (1 mᴍ) for 10 min at 37 °C in homogenates of adult rat whole brain. Changes in brain Na + ,K + -ATPase activity and total antioxidant status (TAS) were studied in the presence of each system separately, with or without Phe, Cys or GSH. TAS value reflects the amount of free radicals and the capacity of the antioxidant enzymes to limit the free radicals in the homogenate. Na + ,K + -ATPase was inhibited by 35-50% and TAS value was decreased by 50-60% by both systems of free radical production. The enzymatic inhibition was completely reversed and TAS value increased by 150-180% when brain homogenates were preincubated with 0.83 mᴍ Cys or GSH. However, this Na + ,K + -ATPase inhibition was not affected by 1.80 mᴍ Phe, which produced a 45-50% increase in TAS value. It is suggested that the antioxidant action of Cys and GSH may be due to the binding of free radicals to sulfhydryl groups of the molecule, so that free radicals cannot induce Na + ,K + -ATPase inhibition. Moreover, Cys and GSH could regulate towards normal values the neural excitability and metabolic energy production, which may be disturbed by free radical action on Na + ,K + -ATPase.

Human serum albumin labeled with technetium-99m was encapsulated together with magnetite particles into phosphatidylcholine/cholesterol liposomes. In order to investigate the stability of this complex and its ability to be used for magnetic drug targeting, the in-vivo distribution after intravenous administration in rats was estimated. For in-vivo targeting an SmCo permanent magnet with intensity ~ 0.3 5 T was attached near the right kidney. Difference between the relative radioactivity in the magnetically targeted right kidney (25.92±5.84%) and non-targeted left kidney (0.93±0.05%) is sufficiently high for relevant clinical applications.

A neural network model with incremental Hebbian learning of afferent and lateral synaptic couplings is proposed,which simulates the activity-dependent self-organization of grating cells in upper layers of striate cortex. These cells, found in areas V1 and V2 of the visual cortex of monkeys, respond vigorously and exclusively to bar gratings of a preferred orientation and periodicity. Response behavior to varying contrast and to an increasing number of bars in the grating show threshold and saturation effects. Their location with respect to the underlying orientation map and their nonlinear response behavior are investigated. The number of emerging grating cells is controlled in the model by the range and strength of the lateral coupling structure.

1-(4-Morpholinomethyl)-tetrahydro-2(1H)-pyrimidinone (mopyridone) exhibited a marked activity against rubella virus (Judith and RA27/3 strains), a MIC 50 value of 0.9 μᴍ and selectivity ratio of 557.7 been found in the case of Judith strain. These data, in addition to the previous ones about its anti-alphavirus effects suggest the compound to be considered as a broad spectrum inhibitor of togavirus replication.

Sixty five methanolic extracts of Sumatran rainforest plants representing 63 species of 21 families were assayed in vivo for antinematodal activity against Bursaphelenchus xylophilus using our cotton ball-fungal mat method. Extracts of 27 plants species from 14 families exhibited antinematodal activity, while 37 species were inactive. Among them, three extracts of Bischofia javanica, Knema hookeriana and Areca catechu exhibited very strong activity at minimum effective dose (MED ) of 0.7 mg/cotton ball (mg/bl.). Eight extracts from Allamanda cathartica, Ervatamia corymbosa, Hoya diversifolia, Bischofia javanica, Derris malacensis, Melastoma melabathricum, Ophiorriza konsteleary and Brucea sumatrana also showed strong activity (MED, 5 mg/bl.).

The activity-guided chromatographic purification of the methanol extract of Knema hookeriana, using pine wood nematodes Bursaphelenchus xylophilus has successfully led to the isolation and characterization of two phenolic antinematodal compounds with minimun effective dose (MED) of 4.5 and 20 μg/cotton ball (μg/ bl.) or 0.018 and 0.073 μᴍ/cotton ball (μᴍ/bl.), respectively. Based on their chemical and spectral properties, these compounds were determined to be 3-undecylphenol (1) and 3-(8Z-tridecenyl)-phenol (2). These compounds were isolated for the first time from this species, and 2 seems to be a novel compound.