Volume 61, Issue 9-10

Needles from 17 different Taxus x media cultivars, belonging to 4 groups showing different growth characteristics, were analyzed using high performance liquid chromatography for their content of 10-deacetylbaccatin III, baccatin III, cephalomannine and paclitaxel (Taxol®). The 4 Taxus x media cultivar groups were: 1.) medium to fast growing and upright form; 2.) slow growing and upright form; 3.) fast growing and spreading form; and 4.) slow growing and spreading form. The purpose of this study was to identify yew cultivars of fast growth rate, upright growth and high taxane content in their needles. The highest content of paclitaxel was found in ‘Coleana’ of group 1 (378 μg/g of the extracted dry weight). Three cultivars in group 1, ‘Coleana’, ‘Stovekenii’ and ‘Hicksii’, make good candidates for taxane extraction because of their high paclitaxel and 10-deacetylbaccatin III content, fast biomass accumulation and upright growing form. They are also good starting materials to develop alternative methods for the production of paclitaxel and its analogous compounds through modern biotechnology approaches.

Nine derivatives of three natural diarylheptanoids, curcumin, demethoxycurcumin and bisdemethoxycurcumin, were prepared. Their antioxidant, free radical scavenging, nitric oxide (NO) inhibitory and cytotoxic activities were evaluated and compared with those of the respective natural compounds. Curcumin (1), demethoxycurcumin (2), demethyldemethoxycurcumin (C3), diacetyldemethoxycurcumin (AC2) and triacetyldemethylcurcumin (AC5) exhibited higher antioxidant activity than quercetin while products from demethylation of 1 and 2 exhibited higher free radical scavenging activity. Compounds AC2 and AC5 were found to be most active in inhibiting breast cancer cells (MCF-7) proliferation with IC 50 values of 6.7 and 3.6 μм, respectively. The activity of AC2 is almost doubled and of AC5 almost tripled as compared to curcumin. Their selectivity towards different cell lines is also more noticeable. Compounds AC2 and AC5 also showed increased activity against a human prostate cancer cell line (DU-145) and non-small lung cancer cell line (NCI-H460) with IC 50 values of 20.4, 16.3 and 18.3, 10.7 μм, respectively.

In the current study, the results of antibacterial, antifungal, and antiviral activity tests of four flavonoid derivatives, scandenone (1), tiliroside (2), quercetin-3,7-O-α-ʟ-dirhamnoside (3), and kaempferol-3,7-O-α-ʟ-dirhamnoside (4), are presented. Antibacterial and antifungal activities of these compounds were tested against Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis, Klebsiella pneumoniae, Acinetobacter baumannii, Staphylococcus aureus, Bacillus subtilis, and Enterococcus faecalis, as well as the fungus Candida albicans by a microdilution method. On the other hand, both DNA virus Herpes simplex (HSV) and RNA virus Parainfluenza-3 (PI-3) were employed for antiviral assessment of the compounds using Madin- Darby bovine kidney and Vero cell lines. According to our data, all of the compounds tested were found to be quite active against S. aureus and E. faecalis with MIC values of 0.5 μg/ml, followed by E. coli (2 μg/ml), K. pneumoniae (4 μg/ml), A. baumannii (8 μg/ml), and B. subtilis (8 μg/ml), while they inhibited C. albicans at 1 μg/ml as potent as ketoconazole. However, only compound 3 displayed an antiviral effect towards PI-3 in the range of 8- 32 μg/ml of inhibitory concentration for cytopathogenic effect (CPE).

Catechol and pyrogallol are allelochemicals which belong to phenolic compounds synthesized in plants. Their antimicrobial activities were investigated on three bacteria (Pseudomonas putida, Pseudomonas pyocyanea, Corynebacterium xerosis) and two fungi (Fusarium oxysporum, Penicillium italicum) phytopathogenic species as test organisms using the disc diffusion method. Both catechol and pyrogallol were found to have antibacterial effects on all the bacteria used in the study at 5 and 10 mм concentrations. Catechol has also been found to have an antifungal effect on the fungi used in the study, whereas no antifungal effects of pyrogallol were observed. The most sensitive species among the bacteria was P. putida which was inhibited by the allelochemicals even at 1 mм concentration.

The cytotoxic potential of three cucurbitacins, 2,3,16,20(R),25-pentahydroxy-11,22-dioxocucurbita- 5-en (cucurbitacin P, 1), 2,3,16,20(R),25-pentahydroxy-22-oxocucurbita-5-en (2) and 2,3,16,20(R),25-pentahydroxy-22-oxocucurbita-5,23(E)-diene (deacetylpicracin, 3), obtained from Cayaponia racemosa was evaluated as their ability to induce brine shrimp lethality, to inhibit the development of sea urchin eggs and tumor cell proliferation, and to lysis mouse erythrocytes. Compounds 1 and 2 were highly toxic with LC 50 of (29.6 ± 9.1) (56.8) and (38.8 ± 3.0) (76.6) μg/mL (μм), respectively, while compound 3 was not effective at the tested concentrations. All tested compounds possessed an inhibitory effect on the proliferation of tumor cell lines, compound 1 being the most active, followed by 2 and 3. Nevertheless, no hemolytic activity or inhibition of the development of sea urchin eggs was observed for these compounds.

Two Brazilian species of Dimerostemma (Asteraceae) were chemically investigated. Two known sesquiterpene lactones (STLs), a germacrolide and an eudesmanolide, were isolated from D. episcopale while D. brasilianum afforded the new germacranolide 1β,5β,10α-trihydroxy- 8α-angeloyloxy-germacra-3,11(13)-dien-6α,12-olide in addition to a known one. Structure identification based on NMR and MS analyses. 1β,10α,4α,5β-Diepoxy-8α-angeloyloxycostunolide isolated from D. brasilianum was studied for its anti-inflammatory activity. This STL completely inhibited DNA binding of the transcription factor NF-κB at a concentration of 5 μм and 10 μм in Jurkat T and Raw 264.7 cells, respectively. Elastase release from human neutrophils was reduced to 50% at a concentrations of 23 μм after stimulation with PAF and of 27 μм after stimulation with fMLP without showing cytotoxic effects. Additionally, elastase was also directly inhibited.

The series of 2,4-dihydroxy-6-methylbenzoates 2-10 (methyl to hexyl orsellinates) prepared by alcoholysis of lecanoric acid (1) - a natural product from the lichen Parmotrema tinctorum (Nyl.) Hale - was submitted to the brine shrimp lethality test (BST), which was also performed for 2,4-dihydroxy-6-methylbenzoic acid (11) (orsellinic acid) and the derivative ethyl-2-hydroxy-4-methoxy-6-methylbenzoate (12) (4-methoxy-ethyl orsellinate), in order to detect new substances with probable antineoplasic activity. Results showed that chain elongation - increase in lipophilicity (log P) - causes a rise in the cytotoxic activity of orsellinates. Hexyl orsellinate (7) showed the highest cytotoxic activity (LC 50 = 31 μм). A correlation between lipophilicity (log P) and cytotoxic activity (log 1/LC 50 ) is presented. Compounds with ramified chains - iso-propyl, sec-butyl and tert-butyl orsellinates (8-10) - were less active than those with the correspondent linear chain. The activities presented by 4-methoxy-ethyl orsellinate (12) and ethyl orsellinate (3) suggest that the hydroxy group at the C-4 position causes effect in the cytotoxic activity of orsellinates against Artemia salina.

Deoxypodophyllotoxin content of the aerial parts of Anthriscus sylvestris Hoffm. growing at different altitudes was evaluated in comparison to the roots. The lignan accumulation in ground parts was at least double compared to aerial ones. In addition antioxidant-guided fractionation of the crude methanol extract of aerial parts was performed with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test. Active fractions contained mainly luteolin-7-O-glucoside and chlorogenic acid. Antioxidant properties of both crude extract and isolated compounds were also investigated with the Briggs-Rauscher (BR) oscillating reaction. A satisfactory agreement between the results obtained with the two methods was observed.

The metabolites of two different Ripartites species, R. tricholoma (A. & S. ex Fr.) Karst. and R. metrodii Huijsm. were investigated. Three new sesquiterpenes were isolated from three different strains. In addition, the strains produced 13-oxo-9(Z),11(E)-octadecadienoic acid, psathyrellon A, 5-desoxyilludosin, an illudane (previously isolated from a Bovista sp.) 96042 and demethylovalicin, five known compounds.

Contents of the hydroxamic acids 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA), and 2,4- dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) in leaves and roots of 14 cultivars of rye, Secale cereale L., were determined. Dynamics of accumulation in three cultivars were evaluated. DIBOA was the main cyclic hydroxamic acid in leaves but the contents differed significantly between the cultivars. Both DIBOA and DIMBOA were present in the roots. Maximum concentration of DIBOA in leaves and DIMBOA in roots was reached between 48-54 h and 54-72 h after germination, respectively. Antifeedant activity of DIBOA towards the aphid Rhopalosiphum padi and the feeding behavior were studied by electronic recording in barley leaves treated with different contents of DIBOA. The deleterious activity of DIBOA could arise by starvation and/or a toxic effect. Additionally, allelopathic potential of pure DIBOA and aqueous extracts of leaves and roots of rye (Tetra-Baer) on the germination of lettuce (Lactuca sativa) and rye (Tetra-Baer) seeds was evaluated. A high percentage of germination inhibition of pure DIBOA and the extracts of leaves and roots was observed. The activity is in agreement with the contents of hydroxamic acids in the plants. The substrates had no allelopathic effect on rye seeds.

The chemical composition of the essential oil obtained from the aerial flowering parts of Ziziphora clinopodioides subsp. bungeana (Juz.) Rech. f. was analyzed by GC and GCMS. Thirty-two components representing 97.1% of the total oil were identified. Oxygenated monoterpenes (94.3%) were the predominant fraction of the oil with pulegone (65.2%), isomenthone (11.9%), 1,8-cineole (7.8%) and piperitenone (6.5%) as the main constituents. Antibacterial activity of the oil and also its two main components (pulegone and 1,8-cineole) were tested against seven bacteria. It was found that the oil exhibited interesting antibacterial activity against Staphylococcus epidermidis, S. aureus, Escherichia coli and Bacillus subtilis with MIC values of 3.75 mg/ml.

The aerial parts of Gontscharovia popovii (B. Fedtsch. and Gontsch.) Boriss. were collected at full flowering stage. The essential oil was isolated by hydrodistillation and analyzed by a combination of capillary GC and GC-MS. Thirty-one components were identified with the main constituent being carvacrol (71.9%), followed by linalool (5.5%), p-cymene (4.5%) and γ-terpinene (4.4%). The in vitro antimicrobial activity of the essential oil of G. popovii was studied against seven Gram-positive and Gram-negative bacteria (Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae) and three fungi (Candida albicans, Saccharomyces cerevisiae and Aspergillus niger). The results of the bioassays showed that the oil exhibited strong antimicrobial activity against all the tested fungi and bacteria except for the resistant bacterium Pseudomonas aeruginosa.

Aristolochia maurorum L. of Jordanian origin has been investigated phytochemically, quantitatively, and biologically. Three atypical alkaloids, namely aristolochic acid I (1), aristolochic acid II (2) and aristolochic acid IIIa (3), have been isolated and identified. Of these known 1-phenanthrenecarboxylic acids, 2 and 3 are reported for the first time from this species. The identified compounds 1-3 were first evaluated biologically as cytotoxic agents against the brine shrimp lethality test (BST), in which compound 1 was found to be the most potent (LC 50 , 4.9 μg/mL). The antiplatelet activity of the methanolic extracts, the acidic fractions of aerial and root parts, and the identified compounds 1-3 were evaluated using an automatic platelet aggregometer and coagulation tracer (APACT 2). Using external reference standards, and a reverse-phase isocratic method, the distribution of aristolochic acid I and aristolochic acid II in different plant parts of Aristolochia maurorum L. during flowering stage was analyzed by PDA-HPLC. A quantitative comparison between two previously reported extraction methods was also made. Roots were found to be the main storage of aristolochic acid I and aristolochic acid II during flowering stage with about 0.22 and 0.108% (w/w), respectively.

It has been previously shown that certain herbicides or plant extracts inhibited the viral infection. The goal of this study was to investigate the effect of Obuda pepper virus (ObPV) infection and herbicide or plant extract treatments on the photosynthetic processes of the host plants to get informations about the interactions of these factors. In Capsicum annuum- ObPV host-virus relations the virus infection slightly increased the activity of photosystem II (PSII), as it was supposed from fluorescence induction parameters. Chlorophyll content of leaves was also elevated probably due to virus-induced growth inhibition. The herbicide Stomp (active ingredient: pendimethalin) incorporated into the soil one week before planting (preplant treatment) together with virus infection even strengthened these effects in agreement with previous observations that this herbicide always did not prevent virus infection or reduce virus concentration in hosts. In ObPV-infected Nicotiana tabacum the structural changes showed similar tendency like in ObPV-infected C. annuum, but PSII efficiency did not significantly differ from that of the control. However, non-photochemical quenching (NPQ) increased because of the strongly decreasing CO 2 fixation activity. Though simultaneous application of a water extract of Cirsium arvense shoot caused a little stronger inhibition of CO 2 fixation, little loss in production was obtained due to significant reduction in virus concentration. In Solanum nigrum-ObPV relation the slightly increasing tendency of the values of actual PSII quantum efficiency could be related to the probably elevated ratio of reaction centre components (increased chlorophyll a/b ratio) in the thylakoids. Application of the herbicide Fusilade S (active ingredient: fluazifop-P-butyl) at 4-6 leaf stage as a postemergence treatment practically prevented systemic virus infection and the virus-induced changes of photosynthesis are probably due to inhibiting the virus infection/replication process.

Stress response capacity (Fv/Fm at 690 nm and F690/F735 at F max ) of untransformed hybrid poplar, Populus × canescens (P. tremula × P. alba), and two transgenic lines overexpressing γ-ECS (γ-glutamylcysteine synthetase) either in the cytosol (cyt-ECS) or in the chloroplast (chl-ECS) was studied in response to the herbicide paraquat (4.0 × 10 -9 to 4.0 × 10 -6 m) for 21 days. Significant differences at sublethal (4.0 × 10 -7 m) and bleaching (4.0 × 10 -6 m) concentrations of paraquat were observed with about a two-fold and eight-fold decrease in the photosynthetic activity (Fv/Fm at 690 nm and F690/F735 at F max ), respectively. None of the gshI transgenic lines (cyt-ECS, chl-ECS) with elevated GSH content exhibited significant tolerance to paraquat. Semiquantitative RT-PCR of the cyt-ECS clone was used for gene expression analysis of the nuclear encoded rbcS gene and the stress responsive gst gene. Expression of the constitutively expressed 26SrRNA ribosomal gene was probed as a control for all RT-PCR reactions. The relative intensities of gene expressions normalized to the level of 26SrRNA intensity showed a 50% decrease in the nuclear encoded rbcS expression and a 120% increase in the stress responsive gst gene expression of the paraquat treated (4.0 × 10 -7 m) samples of the transgenic poplar line (cyt-ECS).

Fifteen wheat genotypes were grown under water deficit to ascertain the role of osmotic adjustment (OA) and the concentration of benzoxazinones in sustaining grain yield. A positive correlation between osmotic adjustment capacity and yield was observed in wheat genotypes cultivated under field conditions. The weight gain of plants exposed to drought was in agreement with the OA values (R 2 = 0.93). However, when wheat plants were infested by cereal aphids, this correlation was not found. The benzoxazinones 2,4-dihydroxy-1,4-benzoxazin- 3-one (DIBOA) and 2,4-dihydroxy-7-methoxy-1,4 benzoxazin-3-one (DIMBOA) are defensive secondary metabolites present in wheat and others cereals. The content of these compounds varied in wheat genotypes and increased with drought and aphid infestation. A positive correlation between weight gain of irrigated-infested plants and drought-infested plants and the contents of benzoxazinones was observed. These results suggest that plants with better OA capacity and high benzoxazinone content should have better field yields.

BOA (2-benzoxazolinone) is a potent phytotoxin present in several graminaceous crops such as rye, maize and wheat. Due to its wide range of phytotoxicity, it is considered as a potential pesticide. A study was conducted to explore the impact of BOA on the radicle and plumule elongation of mung bean (Phaseolus aureus) and associated changes in the macromolecular content - proteins and carbohydrates - and activities of enzymes like amylases, proteases, polyphenol oxidases and peroxidases. BOA significantly reduced the radicle and plumule length of P. aureus, and the contents of proteins and carbohydrates in both root and leaf tissue. On the other hand, activities of hydrolytic enzymes - proteases, amylases, polyphenol oxidases and peroxidases - increased substantially in both root and leaf tissue of P. aureus upon BOA exposure. This indicated that BOA treatment induced stress in P. aureus and enhanced enzyme activities to counter the induced stress and continue the growth. In other words, BOA-induced stress altered the plant biochemical status and related enzyme activities resulting in increased metabolism that serves to provide protection against cellular injury. Such studies providing information about the biomolecular content and enzymatic activities in response to natural products serve as clues for furtherance of knowledge about the modes of action of natural compounds of commercial interest.

Potential biological properties of newly synthesized single and double alkyl chain N-oxides of tertiary amines (NTA) were studied. Individual compounds in each of the series had alkyl chains of different length. Various experiments were performed to determine a mechanism of the interaction between NTA and model and biological membranes. These were measurements of hemolytic efficiencies of NTA (pig erythrocytes), their influence on the transition temperatures (DPPC liposomes), on potassium leakage from cucumber, its growth and chlorophyll content (Cucumis sativus cv. Krak F1), and on the resting membrane potential in alga cells (Nitellopsis obtusa). Also, prevention of erythrocyte membrane lipid oxidation induced by UV irradiation was studied. Potential antioxidative properties of NTA were additionally tested in radical chromogen (ABTS ●+ ) experiments in which antioxidative efficiencies of NTA were compared to that of the standard antioxidant Trolox. It was found that NTA readily interacted with erythrocyte membranes. Their hemolyzing efficiency increased with the alkyl chain length. Slightly more intensive interaction was found for double alkyl chain compounds. Similar results were obtained in DSC experiments, where incorporation of NTA into liposomal membranes shifted the main transition temperatures and caused a broadening of the main transition peaks depending on the alkyl chain length. Double alkyl chain compounds were also found more efficiently influencing the growth of cucumber. Influence of NTA on the resting membrane potential of algae cells was not quite following the alkyl chain length rule found in erythrocyte and liposome experiments. Also potassium leakage and chlorophyll content determined in physiological experiments were not following the increase of lipophilicity of compounds. Most efficiently influencing those parameters were NTA having shorter alkyl chains, and efficiencies of single alkyl chain compounds were evidently stronger. Both methods used to test the antioxidative properties of NTA showed that they depended on the alkyl chain lengths of compounds within each series, but double alkyl chain ones exhibited markedly greater efficiency.

Biosurfactants are surface active substances which reduce interfacial tension and are produced or excreted at the microbial cell surface. We evaluated the biosurfactant production by Aspergillus fumigatus and Phialemonium sp. in solid state processes using fixed-bed column reactors. We evaluated two media, rice husks alone (simple support) and rice husks plus defatted rice bran (complex support), both enriched with either soy oil or diesel oil. The highest water-in-oil emulsifying activity (EA w/o ) obtained was 7.36 EU g -1 produced by A. fumigatus growing on complex support enriched with soy oil and supplied with air at a rate of 60 mL g -1 h -1 , while Phialemonium sp. had a maximum production of 6.11 EU g -1 using the simple support with diesel oil and an aeration rate of 120 mL g -1 h -1 . The highest oilin- water emulsifying activity (EA o/w ) was 12.21 EU g -1 produced by Phialemonium sp. on the complex support enriched with diesel oil and at an aeration rate of 60 mL g -1 h -1 , while A. fumigatus produced a maximum EA o/w of 10.98 EU g -1 when growing on the complex support with no additional carbon source and an aeration rate of 60 mL g -1 h -1 .

Bacillus coagulans has been found to produce several surfactins that are powerful lipopeptide surfactants. Four main components with molecular weights 1007, 1021 and 1035 Da were separated. Their structures have been confirmed by spectrometric and spectroscopic studies and by acid hydrolysis. The compounds were found to represent two pairs of surfactin isoforms in which β-hydroxy-iso-C 14 or anteiso-C 15 fatty acids are linked to the [Leu7] or [Val7] heptapeptide moiety by both an amide group and a lactone bond.

The effect of irradiance during low temperature hardening was studied in a winter wheat variety. Ten-day-old winter wheat plants were cold-hardened at 5 °C for 11 days under light (250 μmol m -2 s -1 ) or dark (20 μmol m -2 s -1 ) conditions. The effectiveness of hardening was significantly lower in the dark, in spite of a slight decrease in the F v /F m chlorophyll fluorescence induction parameter, indicating the occurrence of photoinhibition during the hardening period in the light. Hardening in the light caused a downshift in the far-red induced AG (afterglow) thermoluminescence band. The faster dark re-reduction of P700 + , monitored by 820-nm absorbance, could also be observed in these plants. These results suggest that the induction of cyclic photosynthetic electron flow may also contribute to the advantage of frost hardening under light conditions in wheat plants.

Siderophores play a very important role in the uptake process of iron by bacteria. Due to the so-called active transport the uptake of siderophores by bacteria is very specific, which makes the use of siderophores as effective shuttles for antibiotics in the treatment of infections and other diseases caused by bacteria highly attractive. In order to further investigate the transport and incorporation of siderophores into the bacteria cells, distinct molecular probes are needed. Especially artificial siderophores, that show a specific intrinsic fluorescence, are highly attractive for such monitoring purposes. A promising candidate of such a fluorescent artificial siderophore is bis-2,3-dihydroxybenzoyl-2,6-dimethylamino-pyridine (pyridinochelin, PY). The fluorescence properties of PY were investigated in different solvents and in the presence of different metal ions. It was found that PY in its free form shows a complex fluorescence behavior. In methanol a clear dual fluorescence is observed. In aqueous solution intermolecular interactions with water molecules are determining the intrinsic fluorescence. Upon complexation with metal ions (Me 3+ = Eu 3+ , Tb 3+ , Al 3+ , Fe 3+ ) the fluorescence characteristics changed. The fluorescence quantum yield of PY decreased upon addition of Me 3+ - except for Al 3+ , which showed no fluorescence quenching. The fluorescence decay of PY loaded with metal ions showed a nicely mono-exponential fluorescence decay, which was in contrast to PY in the absence of metal ions. This drastic change in the fluorescence properties of PY upon metal ion complexation makes PY highly attractive as a fluorescence probe for the investigation of siderophore action and siderophore-mediated transport processes

The leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 is the most harmful of the Eucalyptus pests, causing severe losses in wood production through defoliation. Various strategies have been tried and effort spent on the development of methods to control this pest, however no practical and environmentally acceptable one currently exists. In this work the chemical composition of the essential oil of seven Eucalyptus species was identified and the selectivity and sensitivity of antennal receptors of A. sexdens rubropilosa workers to the volatile compounds were determined using the electroantennographic technique (EAG and GC-EAD). Analysis by GC-EAD showed in E. cloesiana and E. maculata, respectively, seventeen and sixteen terpenes that elicited responses in ant workers’ antennae, indicating the potential role of the essential oils as allelochemicals that determine the choice of the foraging material.

We evaluated early and late effects of alloxan-diabetes and subsequent insulin treatment on the kinetic properties of succinate oxidase (SO) in rat liver mitochondria. Diabetic state lowered the SO activity; insulin treatment was effective in restoring the activity only in oneweek diabetic rats. The energies of activation in low and high temperature ranges (E H and E L ) decreased significantly in diabetic animals; once again insulin treatment was partially effective only in the one-week diabetic group. The total phospholipids (TPL) and cholesterol (CHL) contents did not change in one-week groups. In one-month diabetic animals TPL decreased while CHL increased; insulin treatment induced further changes without restoring normality. The lysophospholipid (Lyso), sphingomyelin (SPM), phosphatidylinositol (PI) and phosphatidylserine (PS) content increased in the diabetic state while phosphatidylcholine (PC) and phosphatidylethanolamine (PE) decreased. Insulin treatment had a partial restorative effect. The changes in E H correlated negatively with SPM. The phase transition temperature, T t , decreased in diabetic and insulin-treated groups. These changes correlated positively with the ratios of TPL/PI and TPL/PS. The membrane fluidity decreased in the diabetic state; insulin had a restorative effect only in the one-week group.

In spite of the importance of chemoreception and chemical signals in social organization of lizards, only a few studies have examined the chemical composition of secretions that lizards use for intraspecific communication. The secretion of the femoral glands of male Schreiber’s green lizards (Lacerta schreiberi) contains 51 lipophilic compounds, including several steroids, α-tocopherol, n-C 9 to n-C 22 carboxylic acids and their esteres, and minor components such as alcohols between C 12 and C 24 , two lactones, two ketones, and squalene. These compounds were identified on the basis of mass spectra, obtained by GC-MS. We compared these chemicals with those found in other lizard species, and discussed how environmental conditions could explain the differential presence of chemicals in different lizards. Particularly, the high abundance of α-tocopherol in this lizard is suggested to contribute to avoid oxidation of other lipids in secretions, increasing chemical stability of scent marks in the humid conditions of its habitat.

In this study the enhancement effect of Sophora pachycarpa roots’ acetone extract on the antibacterial activity of gentamycin was evaluated against Staphylococcus aureus. Disc diffusion and broth dilution methods were used to determine the antibacterial activity of gentamycin in the absence and presence of plant extract and its various fractions separated by TLC. A clinical isolate of S. aureus was used as test strain. The active component of the plant extract involved in enhancement of gentamycin’s activity had R ƒ = 0.72 on a TLC plate. The spectral data ( 1 H NMR, 13 C NMR) of this compound revealed that this compound was 5,7,2′,4′-tetrahydroxy-8- lavandulylflavanone (sophoraflavanone G), previously isolated from Sophora exigua. In the presence of 0.03 μg/ mL of sophoraflavanone G the MIC value of gentamycin for S. aureus decreased from 32 to 8 μg/mL (a fourfold decrease). These results signify that the ultra-low concentration of sophoraflavanone G potentiates the antimicrobial action of gentamycin suggesting a possible utilization of this compound in combination therapy against Staphylococcus aureus.