Neutral Lipids of Ruta graveolens L. Grown in vivo and in vitro

The composition of neutral lipids (NL) and fatty acids of acyl-containing components in differentiated tissues ofRuta graveolensL. (Rutaceae) grown in vivo and in vitro is studied. The NL of in vitro cultures differ from NL of the intact plant by a larger content of esters of fatty acids with alcohols, fewer high-molecular-weight alkanes, alkylbenzenes, free alkanols, and linolenic acid     

Effect of polyamines on shoot multiplication and furanocoumarin production in Ruta graveolens cultures

The influence of the polyamines putrescine (Put), spermine (Spr) and spermidine (Spd) on growth and furanocoumarin production was investigated by exogenous addition, at different concentrations, to shoot cultures of Ruta graveolens at different phases of growth. Preliminary studies indicated that addition of Put (20 microM) and Spr (80 microM) had a promotive effect on shoot multiplication rate and number of multiple shoots formed. Spd was toxic, even at lower concentrations. The growth-phase of the culture at the time of exogenous addition of polyamines was found to be an important factor. Put was most effective when added at the lag phase, while Spr was most effective when added in the log phase. Time course studies of growth and furanocoumarin content were carried out for each polyamine and phase of addition. It was seen that maximum production of furanocoumarins (256.8 mg/10 g DW) occurred in the second week when Put was added in the lag phase and 260.5 mg/10 g DW in the fourth week when Spr was added in the log phase. Put addition resulted in a 3.10 fold increase in psoralen, 6.12 in xanthotoxin and 1.46 fold in bergapten production. Spr addition resulted in a 1.31 fold increase in psoralen, 4.11 fold in xanthotoxin and 1.49 fold in bergapten production. Results indicate that alteration of growth and furanocoumarin production kinetics is a combined outcome of choice of polyamine and the phase of culture at the time of exogenous addition. Polyamine addition enabled significant enhancement in production of pharmaceutically important bergapten and xanthotoxin in shoot cultures of Ruta graveolens, which could be explored for commercial production.     

Ruta chalepensis L. (Rutaceae) leaf extract: chemical composition,antioxidant and hypoglicaemic activities

Ruta chalepensis L. (Rutaceae) leaf extract was investigated for its chemical profile and antioxidant and hypoglycaemic properties. The antioxidant effects were investigated by 2,2-diphenyl-1-picrylhydrazyl (DPPH), β-carotene bleaching, and metal chelating activity assays. The carbohydrate-hydrolysing enzymes inhibition assay was used to test the hypoglycaemic potential. R. chalepensis showed a high content of hesperidin and rutin with values of 591.9 and 266.7 mg/g dry extract, respectively. The extract exhibited a promising protection of lipid peroxidation (IC₅₀ value of 16.9 μg/mL) and inhibited both α-amylase and α-glucosidase enzymes in a concentration-dependent manner. The highest activity was found against α-amylase (IC₅₀ value of 69.0 μg/mL). The obtained results support the use of R. chalepensis leaves as healthy food ingredients.     

Studies on the mechanism of aldehyde oxidase and xanthine oxidase

DFT calculations support a concerted mechanism for xanthine oxidase and aldehyde oxidase hydride displacement from the sp(2) carbon of 6-substituted 4-quinazolinones. The variations in transition state structure show that C-O bond formation is nearly complete in the transition state and the transition state changes are anti-Hammond with the C-H and C-O bond lengths being more product-like for the faster reactions. The C-O bond length in the transition state is around 90% formed. However, the C-H bond is only about 80% broken. This leads to a very tetrahedral transition state with an O-C-N angle of 109 degrees. Thus, while the mechanism is concerted, the antibonding orbital of the C-H bond that is broken is not directly attacked by the nucleophile and instead hydride displacement occurs after almost complete tetrahedral transition state formation. In support of this the C=N bond is lengthened in the transition state indicating that attack on the electrophilic carbon occurs by addition to the C=N bond with negative charge increasing on the nitrogen. Differences in experimental reaction rates are accurately reproduced by these calculations and tend to support this mechanism.     

The oxidation of 1-alkyl(aryl)quinolinium chlorides with rabbit liver aldehyde oxidase

1-Alkyl(aryl)quinolinium chlorides are oxidized by rabbit liver aldehyde oxidase at positions C-2 and C-4. The site and the maximum rate of oxidation are dependent on the size and the steric conformation of the N-1 substituent. The presence of a 3-carboxamido group directs the oxidation completely to position C-4, irrespective of the size of the N-substituent. Application of covalent amination in liquid ammonia as an “enzyme model” for the oxidation of these compounds shows little resemblance.     

Effects of interaction of tannins with co-existing substances. VII. Inhibitory effects of tannins and related polyphenols on xanthine oxidase

The inhibitory effects of hydrolyzable tannins, condensed tannins and related polyphenols on the activity of xanthine oxidase (XOD), catalyzing uric acid formation from xanthine, were investigated. Marked differences in the strength of the inhibition were observed. Some of the differences among the monomeric hydrolyzable tannins were due to their molecular weights, reflecting the number of phenolic hydroxyl groups in the molecule. However, the inhibitory activity of several oligomeric hydrolyzable tannins seemed particularly low in spite of their large molecular size. It was also observed that differences in location of acyl groups on the carbohydrate cores caused differences in the inhibitory activity among monomeric and oligomeric hydrolyzable tannins. A caffeic acid derivative (caffeetannin), 3,5-di-O-caffeoylquinic acid (24), also inhibited this enzyme. Galloylation and the degree of polymerization in proanthocyanidins were also shown to affect remarkably the strength of the inhibition. Among the compounds tested in the present study, valoneic acid dilactone (29), isolated from Mallotus japonicus, inhibited the enzyme most effectively. A kinetic study showed that this dilactone inhibited XOD non-competitively. Comparison of the inhibitory effect on XOD, with the binding activity to hemoglobin, for each tannin, suggests that their inhibition of XOD is not based on non-specific binding to the protein. Similar comparison of the inhibitory effect on XOD with the inhibitory effect on the generation of superoxide anion radical (O2-.) from the hypoxanthine-XOD system revealed that the inhibition of O2-. generation by tannins is due to their radical-scavenging activity, and not due to their inhibitory activity upon the enzyme.     

The isolation and purification of diamine oxidase of pea seedlings and pig liver

Enzymes have been extensively used in many industries for the last 20 years. They are becoming more common because of new areas of application. The limitations of applications of enzymes are activity, specificity, stability, and price. Higher enzyme activities and less susceptibility of process conditions are desirable. Therefore, in some cases, purified enzyme extracts are needed. The purpose of this study is the isolation and purification of diamine oxidase (DAO) of pea seedlings and pig liver. The relationship between pea seedlings growth rate and enzyme activity is established. DAO of pea seedlings and pig liver is prepared by way of tissue disruption with homogenization, centrifugation, fractionation with ammonium sulfate, precipitation of inert components, column electrophoresis, and DEAE-cellulose column chromatography. The specific activity of disrupted pea seedlings cells was measured as 0.017 (U/mg protein), and the pig liver DAO activity was measured as 0.00037 (U/mg protein). The specific enzyme activity from pea seedlings was increased to 6.750 (U/mg protein). On the other hand, the specific enzyme activity from pig liver was increased to 0.30 (U/mg protein). The final enzyme extract from pea is 400-fold purer than raw material, and the final enzyme extract from pig liver is 820-fold purer than raw material.     

Chemical composition,phytotoxic and antifungal properties of Ruta chalepensis L. essential oils

The chemical composition, and phytotoxic and antifungal activities of the essential oils isolated by using hydrodistillation from the aerial parts of Tunisian rue were evaluated. Significant variations were observed among harvest periods. The analysis of the chemical composition by gas chromatography/mass spectrometry showed that 2-undecanone (33.4–49.8%), 2-heptanol acetate (13.5–15.4%) and α-pinene (9.8–11.9%) were the main components. The antifungal ability of rue essential oils was tested by using disc agar diffusion against ten plant pathogenic fungi. A high antifungal activity was observed for the essential oil isolated at flowering developmental phase. Furthermore, rue essential oils showed high level of herbicidal activity against several weeds.     

Inhibitory effects of glycyrrhetic acid derivatives on 11 beta- and 3 alpha-hydroxysteroid dehydrogenases of rat liver.

Glycyrrhetic acid (GA), an aglycone of glycyrrhizin (GL), is a potent inhibitor of 11 beta- and 3 alpha-hydroxysteroid dehydrogenases. 11 beta-Hydroxysteroid dehydrogenase activity of rat liver microsomes was potently inhibited by GA, 3-deoxyglycyrrhetic acid (3-deoxyGA), 3-ketoglycyrrhetic acid (3-ketoGA), 3-epiglycyrrhetic acid (3-epiGA) and 11-deoxoglycyrrhetic acid (11-deoxoGA), with I50 values of 2-4 x 10(-7) M. However, 18 alpha-stereoisomers (I50 = 3-7 x 10(-6) M) of GA, 3-deoxyGA and 11-deoxoGA were one tenth less inhibitory on the enzyme activity than the corresponding 18 beta-isomers. On the other hand, 18 alpha-stereoisomers of GA, 3-deoxyGA and 11-deoxoGA inhibited 3 alpha-hydroxysteroid dehydrogenase activity of rat liver cytosol more potently than the corresponding 18 beta-isomers. I50 values of 18 alpha- and 18 beta-isomers were 2 and 7 x 10(-6) M, respectively, in the case of GA, 8 and 20 x 10(-6) M in 3-deoxyGA, 3 and 20 x 10(-6) M in 11-deoxoGA. These results indicate that the 18 beta-conformation of oleanane is important for the inhibition of 11 beta-hydroxysteroid dehydrogenase but on the contrary the 18 alpha-conformation is important for the inhibition of 3 alpha-hydroxysteroid dehydrogenase.     

Inhibitory action of aqueous Ruellia Tuberosa L leaves extract on the corrosion of copper in HCl solution

The inhibitive action of the aqueous extract of Ruellia tuberosa L (ART) on the corrosion of copper in 0.5 ​M HCl was investigated. The inhibition efficiency increased with the extract concentration, acid concentration, as well as increasing the temperature. The Polarization studies revealed that the ART act as a mixed-type inhibitor. Based on the analysis of electrochemical impedance spectroscopy, an equivalent circuit is suggested. The adsorption of the inhibitor ART on the copper surface obeyed the Langmuir adsorption isotherm. From the results of scanning electron microscopy, Fourier transform infrared spectroscopy, and energy-dispersive X-ray spectroscopy the adsorption of ART on the copper surface is confirmed.     

Quantitative study of the structural requirements of phthalazine/quinazoline derivatives for interaction with human liver aldehyde oxidase

Aldehyde oxidase is a molybdenum-containing enzyme distributed throughout the animal kingdom. Although this enzyme is capable of metabolizing a wide range of aldehydes and N-heterocyclic compounds, there is no reported detailed study of physicochemical requirements of the enzyme-substrate interactions. The aim of this study, therefore, was to investigate quantitatively the relationships between the kinetic constants of aldehyde oxidase-catalyzed oxidation of some phthalazine and quinazoline derivatives (as substrates) and their structural parameters. Multiple regression and stepwise regression analyses showed that polarity of phthalazines (expressed as dipole moment mu, cohesive energy density deltaT and an indicator variable for hydrogen-bond acceptor ability of R1 substituent, HBA) had a negative effect on the enzyme activity (leading to the reduction of Vmax and increase of Km). Electron withdrawing substituents in the quinazoline series are favorable for interaction with the enzyme. This finding and also the relationships of 1/Km of phthalazines with the energy of the lowest unoccupied molecular orbital and log Vmax/log Km of phthalazines with degree of bonding of the two nitrogen atoms in the molecules are consistent with the mechanism of action. The reaction involves a nucleophilic attack on an electron-deficient sp2-hybridized carbon atom and formation of an epoxide intermediate following the disruption of the aromatic structure.     

On the oxidation of N-methyl and N-benzylpyrimidin-2- and -4-ones by rabbit liver aldehyde oxidase

The presence of a methyl or benzyl group at N-1 or N-3 of 2- and 4-pyrimidone does not affect the site of oxidation by rabbit liver aldehyde oxidase. From all substrates studies only one product viz. the corresponding N-1 or N-3 substituted uracil has been obtained. The maximum rates of oxidation by free enzyme show an optimum in the pH range 6.5–7.8, which is little influenced by the site and the size of the N-substituent. Application of immobilized enzyme in small scale synthesis gives 1- or 3-R-uracils (R = methyl, benzyl) in 43–78% yield.     

Inhibitory effects of glycyrrhetic acid and its related compounds on 3 alpha-hydroxysteroid dehydrogenase of rat liver cytosol.

Glycyrrhetic acid (GA), aglycone of glycyrrhizin (GL), inhibited potently (I50 = 7 x 10(-6) M) and non-competitively the activity of NAD(P)+-linked 3 alpha-hydroxysteroid dehydrogenase of rat liver cytosol. The inhibition was slightly weaker than that of indomethacin, a potent anti-inflammatory agent, but stronger than that of dexamethasone, another anti-inflammatory agent. GL, GA monoglucuronide, and 3-epi-glycyrrhetic acid also inhibited this enzyme activity, but did so less effectively (I50 = 5-8 x 10(-5) M). Carbenoxolone (GA 3-hemisuccinate) and 3-keto-glycyrrhetic acid showed potent inhibitory effects similar to GA, and 18 alpha-GA showed the most powerful inhibition of the activity.