Band 56, Heft 1-2

The producing strain Streptomyces sp. 1010 was isolated from a shallow sea sediment from the region of Livingston Island, Antarctica. From the culture broth of this strain naturally active secondary metabolites were isolated identical to phthalic acid diethylester (C 12 H 14 0 4 , MW.222); 1, 3-bis (3-phenoxyphenoxy)benzene (C 30 H 22 O 4 , MW.446); hexaned ioic acidd ioctylester (C 22 H 42 O 4 , MW.370) and the new substance 2-amino-9, 13 -dimethylheptad ecanoic acid (C 19 H 39 NO 2 , MW.313). These compounds represent diverse classes of chemical struc­tures and provideevidence for the untapped biosynthetic potential of marine bacteria from Antarctica.

Neodictyoprolenol [(-)-(3 S )-(1,5 Z, 8Z)-undecatrien-3-ol] and dictyoprolenol [ (-) - ( 3 S ) - (1,5 Z , 8Z )-undecadien-3-ol]), which had been proposed as possible biosynthetic intermediates of the sex pheromones of marine brown algae such as dictyopterene B [(-)-trans-1- ((1'E,3' Z )-hexadienyl)-2-vinylcyclopropane], D ' [(+ )-6 -((1' Z )-butenyl)-1,4 -cycloheptadiene] and C' [(+ )-6-butyl-1,4-cycloheptadiene], were again identified in the essential oils from Dictyopteris prolifera, D. latiscula, and in D. undulata, together with the C11-related volatile compounds such as neodictyoprolene, dictyoprolene and dictyopterenes. Incubation of D. prolifela preparation with racemic neodictyoprolenol and dictyoprolenol as substrates showed (S)-enantioselective decreases of the added substrates and increases in dictyopterenes. From these results, a possible pathway to form dictyopterenes is discussed.

Phenolic compounds of purple loosestrife (Lythrum salicaria L.) w ere analysed by the use of liquid chromatography - mass spectrometry (LC/MS) equipped with atmospheric pressure chemical ionisation (APCI) and electrospray ionisation (ESI). The presence of vitexin and orientin as well as their isomers, isovitexin and isoorientin, were confirmed using ion trap multiple stage LC/MS 3 analysis. Several phenolic acids and tannins were also detected. Ellagitannins, vescalagin and pedunculagin, are reported from the plant for the first time.

Alkaloid profiles of two Lupinus species growing naturally in Egypt (L. albus albus [synonym L. term is], L. varius orientalis) in addition to two New World species (L. hartwegii, L. densiflorus) which were cultivated in Egypt were studied by capillary GLC and GLC-mass spectrometry with respect to quinolizidine alkaloids. Altogether 44 quinolizidine, bipiperidyl and proto-indole alkaloids were identified; 29 in L. albus, 13 in L. varius orientalis, 15 in L. hartwegii, 6 in L. densiflorus. Some of these alkaloids were identified for the first time in these plants. The alkaloidal patterns of various plant organs (leaves, flowers, stems, roots, pods and seeds) are documented. Screening for antimicrobial activity of these plant extracts demonstrated substantial activity against Candida albicans, A spergillus flavus and Bacillus subtilis.

Coprinol, a new antibacterial cuparane, was isolated from fermentations of a Coprinus sp. Its biological activities were investigated and its structure was elucidated by spectroscopic methods. The new antibiotic exhibited activitiy against multidrug-resistant Gram -positive bacteria in vitro. Two derivatives were synthesized and their activities compared to the parent compound.

The volatile compounds of a largely consumed Brazilian cashew apple variety (Anacardium occidentale L. var. nanum , Anacardiaceae) were recovered by headspace extraction or simultaneous distillation-extraction. Several compounds including esters (29), terpenes (16), hydrocarbons (9), carboxylic acids (7), aldehydes (7), alcohols (3), ketones (2), lactones (2) and norisoprenoids ( 1 ) were characterized and quantified by gas chromatography-mass spectrometry analyses.

Curcumin, the yellow coloring pigment of turmeric is produced industrially from turmeric oleoresin. The mother liquor after isolation of curcum in from oleoresin known as curcumin removed turmeric oleoresin (CRTO) was extracted three times with n-hexane at room temperature for 30 min to obtain turmeric oil. The turmeric oil was subjected to fractional distillation under vacuum to get two fractions. These fractions were tested for antifugal activity against A spergillu s flavus, A . parasiticus, Fusarium moniliform e and Penicillium digitatum by spore germination method. Fraction II was found to be m ore active. The chemical constituents of turmeric oil, fraction I and fraction II were determined by GC and identified by GCMS. Aromatic turmerone, turmerone and curlone were major compounds present in fraction II along with other oxygenated com pounds.

The cytotoxic activity of two pentacyclic triterpenoids (ursolic acid and α-amyrine) isolated from the methanolic extract of the aerial parts from Erica andevalensis, whose structures have been established on the basis of spectroscopic and chemical evidence, has been assessed against three human cancer cell lines, TK -10 (renal adenocarcinom a), MCF-7 (breast adenocarcinoma) and UACC-62 (melanoma), recommended by NCI (National Cancer Institute) and we also evaluated the antimitotic effect in root meristematic cells of Allium cepa. Ursolic acid was found to possess the highest cytotoxic activity.

The antimicrobial activity of fifteen semisynthetic labdane-type diterpenes derived from the two major natural compounds 3 and 4 of the resin "ladano" of Cistus creticus is reported. The chloroethyl carbamidic esters 15 and 20 showed the strongest antimicrobial activity against Gram (+), Gram (-) bacteria and pathogenic fungi.

The effect of minimum inhibitory concentrations (MICs) of six novel fenpropimorph deri­vatives on lipid and sterol composition of Candida albicans, Cryptococcus neoformans, Malassezia pachydermatis and Malassezia furfur was investigated. The MICs for the most effective derivatives were found in the range from 3.7 to 56.7 μм and were 2 -3 times lower compared to the commercial fungicide bifonazol. The more efficient fenpropimorph derivatives were the piperidine derivative for C. albicans and the allylamine derivative for Cr. neoformans, M. pachydermatis and M. furfur . The inhibitor in the growth medium reduced the unsaturation index of the total lipid content in M. furfur and C.

In the course of our continuing work on transformation of monoterpenes by microorga­nisms, the biotransformation of (+)-and(-)-carvone was carried out. The metabolites formed during microbial transformations were screened using a simple, rapid and efficient technique: Headspace-solid phase microextraction the application of this technique are described.

Elicitation of cultured chickpea (Cicer arietinum L.) cells stimulates a signal transduction pathway leading to several rapid responses: (1) oxidative burst, (2) extracellular alkalinisation, (3) extracellular acidification, (4) transient K + efflux, and (5) activation of defence related genes all within 2 hours. Induced genes are encoding acidic and basic chitinases, a thaumatin-like protein and isoflavone reductase. All these elicitor-induced responses are in­ hibited by the Ser/Thr protein kinase inhibitor staurosporine and the anion channel blocker anthracene-9-carboxylic acid but stimulated by the Ser/Thr protein phosphatase 2A inhibitor cantharidin. The oxidative burst leads to a transient extracellular H 2 O 2 accumulation which seems to be preceded by O 2 - production, indicating dismutation of O 2 - to H 2 O 2 . The oxidative burst is accompanied by transient alkalinisation of the culture medium which is followed by long-lasting extracellular acidification. An 80 percent inhibition of the alkalinisation after complete inhibition of the H 2 O 2 burst with diphenylene iodonium indicates that the elicitor induced increase of extracellular pH is mainly based on a proton consumption for O 2 - dismutation. A simultaneous deactivation of the plasma membrane H + -ATPase during oxidative burst and extracellular alkalinisation is also suggested. The elicitor-stimulated extracellular acidification is inhibited by the plasma membrane H + -ATPase inhibitor N, N'-dicyclohex-ylcarbodiimide assuming a reactivation of the H + -ATPase 25 min after elicitation. Extracellular acidification seems not to be necessary for elicitor-induced activation of defence related genes. Opposite modulation of K + and proton fluxes after elicitation and/or treatment with the H + -ATPase effectors fusicoccin or N, N'-dicyclohexylcarbodiimide indicate that the elicitor induced transient K + efflux is regulated by a K + /H + exchange reaction.

In the present study the leaf movement rhythm was perturbed by the application of DC current pulses 15 μA, 10 seconds, voltage applied: 10 V) to the upper part of the pulvinus, passing through the pulvinus and its stalk. The pulses were applied at four different positions of the leaflets: when the leaves were at the lowermost position, when moving up, at the uppermost position and when moving down. The pre-perturbed and the post-perturbed rhythms were compared. We found that the rhythms were shifted in phase and the phase shifts observed at the four different positions of the leaflets were significantly different in magnitude as well as direction. Furthermore, we could also observe phase advances, which is in contrast to an earlier finding. A phase response curve (PRC) was constructed to illustrate the sensitivity of the oscillating leaflet system to DC pulses. Substantial delays of about 50 s (as compared to the period of about 200 s) were obtained when pulses were administered at the lowermost position and when leaflet were moving upwards, while advances or no phase shifts were recorded in the uppermost position and when leaflet were moving down respectively.

The antimicrobial activity of four propolis samples collected from Upper Egypt against Staphylococcus aureus; Escherichia coli and Candida albicans was evaluated. There was a variation in the antimicrobial activity according to the propolis origin. Banisweif propolis showed the highest antimicrobial activity against Staphylococcus aureus, Escherichia coli and Candida albicans, but Fayoum propolis had moderate activity against all tested pathogens. Propolis collected from Assiut and Souhag gave lower antimicrobial activity. Propolis samples were investigated by GC/MS, 71 compounds were identified, 14 being new for propolis. Banisweif propolis is characterized by the presence of 7 caffeate esters and 4 triterpenoids. Fayoum propolis showed the highest amount of lactic acid and the presence of 3 chalcones. But Assiut propolis is characterized by the presence of 4 prenylated coumar-ates. Souhag propolis is characterized by the presence of 5 aliphatic dicarboxylic acids and some other new compounds to propolis.

Scolus secretions and hemolymph of caterpillars of Satumia pyri fed with two different foodplants (Crataegus monogyna, Prunus spinosa) were chemically analyzed and their chemical similarities determined. The secondary-compound patterns obtained for the two body fluids showed no significant differences when compared between the two groups of alterna­ tively fed last-instar larvae. Thus, the composition of these fluids of full-grown caterpillars is not influenced by the larval diet. However, younger larvae on P. spinosa revealed a diversity of compounds differing significantly from that of larger caterpillars fed with either C. mono­gyna (both body fluids) or P. spinosa (hemolymph only). This indicates that, on the one hand, the hemolymph composition is adapted to the changing physiological requirements of the given instars whereas, on the other hand, the defensive mixtures remain unaltered in the late larval instars due to a constant spectrum of potential enemies.

Argentatins, Insect Growth Regulators, Acetylcholinesterase The methanolic extract from aerial parts of Parthenium argentatum, afforded argentatin A and B. These compounds were evaluated for their effect on the fall armyworm (Spodoptera frugiperda). Toosendanin, a commercial insecticide derived from Melia azedarach was used as positive control. When tested for activity, using neonate larvae into the no-choice artificial diet bioassays, argentatin A, argentatin B and methanol extract caused significant growth inhibitory activity with GC 50 of 17.8, 36.1 and 6.4 ppm at 7 days, respectively, and increased the development time of surviving larvae in a concentration-dependent manner with RGI values of 0.40, 0.60 and 0.26, at 25.0, 25.0 and 5.0 ppm. respectively. In addition, it was possible to observe in most of the treated groups a significant delay in the time of pupation, adult emergence and deformities. Acute toxicity against adults of S. frugiperda was also found, MeOH extract had the most potent activity with LD 50 value of 3.10 ppm. In addition, MeOH extract and argentatin A caused acetylcholinesterase inhibition of 93.7% and 90.0%, at 5.0 and 50.0 ppm, respectively; whereas argentatin B had only slight inhibitory activity. Therefore, the MeOH extract was identified as insecticidal extract from P. argentatum with activity at concentrations above 15.0 ppm.

Mariannaeapyrone ((E)-2-(1,3,5,7-tetramethyl-5-nonenyl)-3,5-dimethyl-6-hydroxy-4H-pyran-4-one) is a new fungal metabolite isolated from fermentations of the common mycophilic deuteromycete Mariannaea elegans. The chemical structure of the 4-pyrone was determined by spectroscopic techniques. Mariannaeapyrone is a selective inhibitor of the thromboxane A2 induced aggregation of human platelets, whereas only weak cytotoxic and antimicrobial effects could be observed.

The CCl 4 -induced development of liver damage was studied in monolayer cultures of pri­mary rat hepatocytes: (1) CCl 4 caused accumulation of triglycerides in hepatocytes following cytochrome P450 induction with β-naphthoflavone or metyrapone. Ethanol or a high dose of insulin plus triio­dothyronine had the same effect. (2) CCl 4 increased the synthesis of fatty acids and triglycer­ ides and the rate of lipid esterification. Cholesterol and phospholipid synthesis from acetate was also increased. (3) CCl 4 reduced β-oxidation of fatty acids as assessed by CO 2 -release and ketone body formation. Hydrolysis of triglycerides was also reduced. (4) The content of unsaturated fatty acids in microsomal lipids was decreased by almost 50% after incubation with CCl 4 , while saturated fatty acids increased slightly. (5) CCl 4 exerted a pronounced inhibitory effect on the exocytosis of macromolecules (albumin), but did not affect secretion of bile acids from hepatocytes.

Haptoglobin is one of acute phase glycoproteins often used as markers in glycopathology studies. In this work the oligosaccharide structures of haptoglobin from 'healthy' subjects have been studied in detail, taking into consideration the possible dependence of glycosylation on the phenotype. About 75% of charged haptoglobin glycans were of biantennary complex structure, and some of them lacked one terminal sialic acid molecule. Triantennary structures made up almost 25% of the charged glycans pool, and highly branched tetrasialy-lated oligosaccharides did not exceed 1%. The main difference between haptoglobin derived from the sample of pooled 44 sera and from the 2 -2 phenotype individual concerned the relative content of trisialylated oligosaccharide with one 2 -3 linked sialic acid residue. The oligosaccharide profile of haptoglobin derived from serum of a patient suffering from con­ genital disorder of glycosylation was compared to 'healthy' controls. It was shown, that four main glycans are identical in patient and 'normal' haptoglobins. Some alterations were found in the relative content of mono-, bi-, and trisialylated glycans as well as in the appearance of some tracely abundant oligosaccharides in haptoglobin of the patient with congenital dis­ order of glycosylation.

The effect of different L-phenylalanine (Phe) concentrations (0.12-12.1 mм) on acetylcho­ linesterase (AChE), (Na + ,K + )-ATPase and Mg 2+ -ATPase activities was investigated in homogenates of adult rat whole brain and frontal cortex at 37 °C. AChE, (Na + ,K + )-ATPase and Mg 2+ -ATPase activities were determined after preincubation with Phe. AChE activity in both tissues showed a decrease up to 18% (p<0.01) with Phe. Whole brain Na + ,K + -ATPase was stimulated by 30-35% (p<0.01) with high Phe concentrations, while frontal cortex Na + ,K + -ATPase was stimulated by 50-55% (p<0.00l). Mg 2+ -ATPase activity was increased only in frontal cortex with high Phe concentrations. It is suggested that: a) The inhibitory effect of Phe on brain AChE is not influenced by developmental factors, while the stimulation of Phe on brain Na + ,K + -ATPase is indeed affected; b) The stimulatory effect of Phe on rat whole brain Na + ,K + -ATPase is decreased with age; c) Na + ,K + -ATPase is selectively more stimulated by high Phe concentrations in frontal cortex than in whole brain homogenate; d) High (toxic) Phe concentrations can affect Mg 2+ -ATPase activity in frontal cortex, but not in whole brain, thus modulating the amount of intracellular Mg 2+ .

The binuclear zinc(II) complex, [Zn 2 (HPTP)(CH 3 COO)] 2+ was found highly active to cleave DNA (double-strand super-coiled DNA, pBR322 and φχ174) in the presence of hy­drogen peroxide. However, no TBARS (2-thiobarbituric acid reactive substance) formation was detected in a solution containing 2 -deoxyribose (or 2′-deoxyguanosine, etc); where (HPTP) represents N,N,N′-N′-tetrakis(2-pyridylmethyl)-1,3-diamino-2-propanol. These facts imply that DNA cleavage reaction by the binuclear Zn(II)/H 2 O 2 system should be due to a hydrolytic mechanism, which may be attributed to the enhanced nucleophilicity but depressed electrophilicity of the peroxide ion coordinated to the zinc(II) ion. DFT (density-functional theory) calculations on the peroxide adduct of monomeric zinc(II) have supported the above consideration. Similar DFT calculations on the peroxide adducts of the Al(III) and La(III) compounds have revealed that electrophilicity of the peroxide ion in these com­ pounds is strongly reduced. This gives an important information to elucidate the fact that La 3+ can enhance the growth of plants under certain conditions.

DFT calculations were done for the (hydroperoxo)metal complexes with η 1 -coordination mode, where metal ions are Fe(III), Al(III), Cu(II) and Zn(II). Results shows that 1) the electron density at the two oxygen atoms of the hydroperoxide ion is highly dependent on the angle O-O-H in M-OOH species and the difference in electron density between the two oxygen atoms reaches a maximum at the angle O-O-H = 180°, 2) total electron density at the two oxygen atoms of the peroxide ion increases by approach of methane to the (hydro-peroxo)metal species in the cases of Fe(III) and Cu(II); on the other hand, significant decrease of the electron density on peroxide oxygen atoms was observed for the cases of Al(III) and Zn(II) compounds. These findings suggest that the (hydroperoxo)metal species acts as an electrophile in the former cases (M = Fe(III), Cu(II)) and as a nucleophile for the latter two compounds (M = Zn(II), Al(III)). The electrophilicity observed for the Fe(III) and Cu(II) complexes is attributed to the presence of unoccupied-or half-filled d-orbitals interacting with the hydroperoxide ion. 3) Two oxygen atoms of the (hydroperoxo)-com-pounds of Fe(III) and Cu(II) complexes exhibit quite different reactivity toward the substrate, such as methane. When methane approaches the oxygen atom which is coordinated to a metal ion, a strong decrease of electron density at the methane carbon atom occurs with concomitant increase of electron density at the peroxide oxygen atoms inducing its hetero-lytic O-O cleavage. When methane approaches the terminal oxygen atom, an oxidative coupling reaction occurs between peroxide ion and methane; at first a nucleophilic attack by the terminal electron-rich oxygen atom occurs at the carbon atom to induce C-O bond formation, and a subsequent oxidative electron transfer proceeds from substrate to the metal-peroxide species yielding CH 3 -OOH, CH 3 OH, or other oxidized products. These results clearly de­monstrate that the (hydroperoxo)-metal compound itself is a rather stable compound, and activation of the peroxide ion is induced by interaction with the substrate, and the products obtained by the oxygenation reaction are dependent on the chemical property of the sub­ strate, redox property of a metal ion, and stability of the compounds formed in the intermediate process.

The micellization process of dodecyltrimethylammonium chloride (DTAC) and bromide (DTAB) was studied at 313 K. Nuclear magnetic resonance and calorimetric methods were used. The calorimetric titration curves permitted determination of the critical micelle concentration (CMC) and enthalpy of the micellization process (ΔH m ) of the compounds studied. The results obtained were compared to those obtained at 298 K. It was found that calorimetric curves obtained at 313 K for both compounds were similar to each other in contrast to 298 K. Especially a great difference in the shape of curves was observed for DTAC. NMR ( 1 H NMR and 13 C NMR) spectra were taken below and above the CMC values and chemical shifts (6) analysed as a function of concentration of the compounds. Comparison of chemical shift-concentration plots with those obtained from measurements performed at lower temperature showed that chemical shifts are of very similar character in both cases for analyzed groups. However, there are some quantitative differences that indicate at smaller difference in hydration of DTAB and DTAC micelles at elevated temperature. This may be the reason of decrease of differences between micellization processes of DTAC and DTAB compounds. The smaller hydration may be, in turn, the result of diminishing differences in physicochemical properties of bromide and chloride ions with temperature.

This work describes a comparative qualitative and quantitative chemical analysis of Maytenus ilicifolia and Maytenus robusta (Celastraceae), extracts by high-resolution gas chromatography (HRGC), using external standards as the method of determination and thin layer chromatographic (TLC). The results show that both plants have a similar chromatographic profile. However, M. robusta exhibited about three times higher concen tration of triterpene friedelin than M. ilicifolia.