Volume 66, Issue 5-6

In order to find new lead compounds with antitumour activies, thirteen new fatty acid esters of 20(S)-protopanaxadiol (PPD) were synthesized using oleoyl chloride or fatty acids and N,N’-dicyclohexylcarbodiimide (DCC). Their cytotoxic activities were tested using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] method, and the structure- activity relationships between the fatty acid esters of PPD and their cytotoxic activities are discussed.

From the aerial parts of Dendrobium secundum, a new bibenzyl, namely 4,5,4’-trihydroxy- 3,3’-dimethoxybibenzyl, was isolated, along with four known compounds which included brittonin A, moscatilin, syringaresinol, and ferulic acid. All of these isolates showed appreciable DPPH free radical scavenging potential, except for brittonin A, which was devoid of activity.

In the course of searching for new whitening agents, we have found that the methanol extract of dried skin of Allium cepa shows potent melanin biosynthesis inhibitory activity in B16 melanoma cells. Bioassay-guided fractionation led to the isolation of quercetin-3’-O-β- D-glucoside (1) from the methanol extract of dried skin of A. cepa, which inhibited melanin formation in B16 melanoma cells with an IC 50 value of 38.8 μM and mushroom tyrosinase with an IC 50 value of 6.5 μM using L-tyrosine and 48.5 μM using L-dihydroxyphenylalanine as substrates, respectively. In addition, the antioxidant activity of 1 was evaluated in the oxygen radical absorbance capacity assay; it showed 3.04 μmol Trolox equivalents/mmol. 1 was shown to be a promising ingredient that could be useful for treating hyperpigmentation and for protecting against oxidative stress.

Phytoecdysteroids from aerial parts of Silene guntensis B. Fedtsch were investigated and three phytoecdysteroids were isolated: 2,3-diacetate-22-benzoate-20-hydroxyecdysone (1), 2-deoxy-20-hydroxyecdysone (2), and 20-hydroxyecdysone (3). Their chemical structures were elucidated by DEPT, COSY, 1 H and 13 C NMR spectroscopy. The isolated compounds 1 - 3 and crude extracts were evaluated for their antiproliferative and antioxidant activities. They exhibited substantial inhibition of cell growth against human cervix adenocarcinoma (HeLa), hepatocellular carcinoma (HepG-2), and breast adenocarcinoma (MCF-7) cells. The chloroform extract showed potent cytotoxic effects [IC 50 values (26.58 ± 1.88) μg/mL, (20.99 ± 1.64) μg/mL, and (18.89 ± 2.36) μg/mL, respectively]. The new compound 1 was mildly cytotoxic compared to extracts [(127.97 ± 11.34), (106.76 ± 7.81), and (203.10 ± 19.56) μg/mL, respectively]. Water and n-butanol extracts exhibited good antioxidant activities [IC 50 values of (68.90 ± 6.45) μg/mL and (69.12 ± 5.85) μg/mL, respectively].

Five oxindole alkaloids, three plumerane-type alkaloids, subtype haplophitine, and one aspidospermatane-type alkaloid, subtype tubotaiwine, were isolated from the medicinal plants Aspidosperma rigidum and A. schultesii. One compound was identified as the transoid conformer of 18-oxo-O-methylaspidoalbine which was not previously described. The antiparasitic activity of all compounds against Trypanosoma cruzi and Leishmania infantum and their non-specific cytotoxicity against mammalian cells were also determined.

Cycloartenyl stearate (1a), lupenyl stearate (1b), sitosteryl stearate (1c), and 24-methylenecycloartanyl stearate (1d) (sample 1) from the air-dried leaves of Syzygium samarangense exhibited potent analgesic and anti-inflammatory activities at effective doses of 6.25 mg/kg body weight and 12.5 mg/kg body weight, respectively. Sample 1 also exhibited negligible toxicity on zebrafish embryonic tissues. There were incidences of mortality upon direct exposure of sample 1 to dechorionated embryos, but higher mortality and aberration were observed during intact chorion treatment.

The activities of two triterpenoids, ocotillone and cabraleadiol, and four limonoids, methyl angolensate, 3-β-deacetylfi ssinolide, 7-deacetoxy-7-oxogedunin, and β-photogedunin, isolated from arillus of Carapa guianensis and fruits and seeds of Cabralea canjerana (Meliaceae), were evaluated against the fall armyworm Spodoptera frugiperda. Gedunin was used as a positive control. 7-Deacetoxy-7-oxogedunin and β-photogedunin reduced the pupal weight as occurred with gedunin. Cabraleadiol, 3-β-deacetylfi ssinolide, and 7-deacetoxy-7-oxogedunin prolonged the larval phase similar to the control (gedunin) of approximately 1.2 days at 50.0 mg kg -1 . The highest insecticidal activity was obtained for β-photogedunin.

A bioactivity-guided fractionation of the ethyl acetate fraction of the flowers of Onopordum alexandrinum L. (Asteraceae) yielded a new flavonoidal glycoside designated as acacetin-7-O-galacturonide (9), alongside with nine known flavonoids; 6-methoxy-apigenin (hispidulin) (1), acacetin (2), apigenin (3), luteolin (4), kaempferol (5), eriodictyol (6), apigenin- 7-O-glucoside (7), luteolin-7-O-glucoside (8), and kaempferol-3-O-rutinoside (10). The compounds were assayed for their hepatoprotective activity against CCl 4 -induced hepatic cell damage in rats and free radical scavenging activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH). Compounds 4, 6, 9, and 10 have not been previously reported from flowers of O. alexandrinum L., and this is the first report of acacetin-7-O-galacturonide (9) in nature which has also shown significant hepatoprotective and free radical scavenging effects. The isolated compounds were identified using different spectroscopic methods (UV, 1 H NMR, 13 C NMR, HMQC, HMBC, and COSY).

Citronellol, an oxygenated monoterpene, is found naturally in the essential oils of several aromatic plants and has been reported to exhibit growth inhibitory and pesticidal activities. However, its mechanism of action is largely unexplored. We investigated the effect of citronellol, which is lipophilic in nature on membrane integrity in terms of lipid peroxidation, conjugated dienes content, membrane permeability, cell death, and activity of the enzyme lipoxygenase in roots of hydroponically grown wheat. Citronellol (50 - 250 μM) caused a significant inhibition of root and shoot growth. Furthermore, exposure to citronellol enhanced the solute leakage, increased the malondialdehyde content and lipoxygenase activity, and decreased the conjugated diene content. This indicates that citronellol induces generation of reactive oxygen species (ROS) resulting in lipid peroxidation and membrane damage. This was confirmed by in situ histochemical studies indicating cell death and disruption of membrane integrity. We conclude from this study that citronellol inhibits the root growth by ROS-mediated membrane disruption.

Fatty acids of twelve species of cyanobacteria grown under different photoautotrophic conditions were studied and their composition was compared with literature data of many other species. We have come to the conclusion that the lipids of cyanobacteria do not contain fatty acids with a chain longer than 18 carbon atoms. In our opinion, omission of an analytical procedure, i.e. purification of fatty acid methyl esters before gas chromatography, leads to incorrect interpretation of the results. Absence or presence of fatty acids was suggested as a useful taxonomic marker and a proper diagnostic indicator in the commercial application of cyanobacterial biomass.

The inclusion complex of hydroxypropyl-β-cyclodextrin (HBβCD) and phytosterols (PSs) was prepared and characterized by thermogravimetric analysis (TGA) and infrared (IR) spectroscopy. Biotransformation of the inclusion complex of phytosterols and hydroxypropyl- β-cyclodextrin (PSs-HBβCD) by Mycobacterium neoaurum to 1,4-androstadiene-3,17-dione and 4-androstene-3,17-dione [AD(D)] was studied. The TGA and IR results indicated that the thermal stability of PSs was improved in the complex with HBβCD. Biotransformation improved the solubility of PSs in the aqueous medium a lot because the AD(D) production was increased remarkably compared with the control, but growth of the bacteria was inhibited in the presence of HBβCD. The optimal inclusion ratio, ultrasonic treating time, dosage, and time of addition of PSs-HBβCD complexe were found to be 2:1, 10 min, 1.5 g/30 ml medium, and 48 h after incubation, respectively. This inclusion technique not only increased the availability of the substrates for the microorganisms, but also the capability of these microorganisms to produce AD(D) from PSs.

This report gives an insight into the specific changes in the transcription of four key carotenogenic genes [encoding geranylgeranyl diphosphate synthase (crtE), phytoene desaturase (crtI), phytoene synthase lycopene cyclase (crtYB), and astaxanthin synthase (ast), respectively] in Xanthophyllomyces dendrorhous cultures, with regard to dissolved oxygen (DO) contents of 10%, 25%, and 40% air saturation, respectively. 25% DO proved to be the most beneficial for yeast growth, transcription of carotenogenic genes, and astaxanthin content.

The aim of the present study was to purify and characterize a toxic protease secreted by the pathogenic Photobacterium damselae subsp. piscicida strain CP1 originally isolated from diseased cobia (Rachycentron canadum). The toxin isolated by anion exchange chromatography, was a metalloprotease, inhibited by L-cysteine, ethylenediaminetetraacetic acid (EDTA), ethylene glycol-bis(β-aminoethyl ether)N,N,N’,N’-tetraacetic acid (EGTA), 1,10-phenanthroline, N-tosyl-L-phenylalanine-chloromethyl ketone (TPCK), and N-α-ptosyl- L-lysine-chloromethyl ketone (TLCK), and showed maximal activity at pH 6.0 - 8.0 and an apparent molecular mass of about 34.3 kDa. The toxin was also completely inhibited by HgCl 2 , and partially by sodium dodecyl sulfate (SDS) and CuCl 2 . The extracellular products and the partially purified protease were lethal to cobia with LD 50 values of 1.26 and 6.8 μg protein/g body weight, respectively. The addition of EDTA completely inhibited the lethal toxicity of the purified protease, indicating that this metalloprotease was a lethal toxin produced by the bacterium.

This study was aimed at characterizing allelic variations of erythrocyte histone H1.b by comparing the electrophoretic patterns of histone H1.b from individuals of grey partridge (Perdix perdix) population. As two alloforms, H1.b1 and H1.b2, were discerned in the screening gels, the histone H1.b was regarded to be a polymorphic protein encoded by a gene with two codominant alleles, b 1 and b 2 , at a locus. The tested population was found to be at Hardy-Weinberg equilibrium (χ2 = 0.834, p = 0.361), with only a minor heterozygote deficiency (fixation index F = 0.136). Since the histone H1.b alloforms were identified in a two-dimensional gel containing sodium dodecyl sulfate, with no significant differences in their migration pattern in an one-dimensional acetic acid polyacrylamide gel, we assumed that the H1.b alloforms possessed a similar net charge and differed in their apparent molecular weights. A comparison of N-bromosuccinimide-cleaved and α-chymotrypsin-digested products of histone H1.b alloforms revealed slight differences in the velocity of C-terminal peptides and a similarity in migration of their N-terminal peptides in one-dimensional sodium dodecyl sulfate-polyacrylamide gel. Therefore, it seemed that the histone H1.b alloforms might differ in this amino acid sequence in a protein segment between N-bromosuccinimide cleavage site and the very C-terminus.

Api SI and Api SII are serine proteases of the honeybee venom containing allergenic determinants. Each protease consists of two structural modules: an N-terminal CUB (Api SI) or a clip domain (Api SII) and a C-terminal serine protease-like (SPL) domain. Both domains are connected with a linker peptide. The knowledge about the structure and function of Api SI and Api SII is limited mainly to their amino acid sequences. We constructed 3-D models of the two proteases using their amino acid sequences and crystallographic coordinates of related proteins. The models of the SPL domains were built using the structure of the prophenoloxidase-activating factor (PPAF)-II as a template. For modelling of the Api SI CUB domain the coordinates of porcine spermadhesin PSP-I were used. The models revealed the catalytic and substrate-binding sites and the negatively charged residue responsible for the trypsin-like activity. IgE-binding and antigenic sites in the two allergens were predicted using the models and programs based on the structure of known epitopes. Api SI and Api SII show structural and functional similarity to the members of the PPAF-II family. Most probably, they are part of the defence system of Apis mellifera

Coal is the most abundant of the fossil fuels, with reserves estimated at 102 billions of tons. The feasibility of using coal as a fuel depends upon reducing emissions of gas when it is burnt, such as carbon dioxide (CO 2 ), sulfur oxides (SO x ), and nitrogen oxides (NO x ). The removal of CO 2 with microalgae may be one of the most efficient ways of reducing this gas, without the need for radical changes in the world’s energy supply and production methods. Spirulina sp. LEB-18 and Scenedesmus obliquus LEB-22 were cultivated in serial tubular photobioreactors, with the aim of measuring the potential of CO 2 biofixation and the resistance of the microalgae to SO 2 and NO. Spirulina sp. and S. obliquus had CO 2 biofixation scores of 0.27 and 0.22 g L-1 d-1, respectively. Both microalgae were resistant to SO 2 and NO, and grew during the 15 d they were cultivated, which proves that using microalgae is an efficient method of biofixation of CO 2 emitted when fossil fuels are burnt.