Comparative study on phenolic content and antioxidant activity during maturation of the olive cultivar Chemlali from Tunisia

For the first time the identification and quantification of phenolic compounds of the Olea europaea L. cv. Chemlali olive were carried out to examine their profile during maturation. The phenolic composition was studied by using reverse-phase high-performance liquid chromatography during all steps of fruit development. Oleuropein is the abundant phenolic compound in Chemlali olive, and its concentration increases during maturation. An indirect relationship between oleuropein content in olive fruit and hydroxytyrosol was observed. Weak changes in the amounts of the other phenolic monomers and flavonoids were also observed. The total phenolic content varied from 6 to 16 g/kg expressed as pyrogallol equivalents. Its highest level was found at the last maturation period. The antioxidant capacity of olive extracts was evaluated by measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl. The IC(50) values of the olive extract ranged from 3.2 to 1.5 microg/mL. There was a correlation between antioxidant activity and total phenolic content of samples. The antioxidant activity increased with maturation. This could be attributed to the increase of the tolal phenol level with fruit development.     

Structure-physicochemical function relationships of soybean beta-conglycinin heterotrimers

We purified four single molecular species of beta-conglycinin heterotrimers consisting of the alpha and beta subunits or the alpha' and beta subunits from mutant soybean cultivars lacking the alpha or alpha' subunit, respectively, and examined their structural features and physicochemical functions. The extent of the hydrophobicities of the heterotrimers was related to the number of the alpha or alpha' subunit. The thermal stabilities of the heterotrimers were mainly conferred by the subunit which had lower thermal stability. Solubilities at low ionic strength (mu = 0.08) of the heterotrimers containing the alpha or alpha' subunit were very similar to those of the alpha and alpha' homotrimers, respectively. Emulsifying abilities and heat-induced associations of the heterotrimers containing one beta subunit were similar to those of the alpha or alpha' homotrimer, whereas those of the heterotrimers containing two beta subunits were similar to those of the beta homotrimer.     

Nutrient Stress,Ecophysiological, and Metabolic Aspects of Olive Tree Cultivars

This research, aimed at evaluating stress on the olive tree, utilized cultivars ‘Meski’ and ‘Chetoui’ in regards to the removal of major elements such as nitrogen (N), phosphorus (P), magnesium (Mg), and potassium (K). The experiment was conducted under greenhouse conditions in hydroponics culture. Measurements of leaf area showed a non-substantial and weak effect due to phosphorus, potassium, or magnesium deficiency compared to nitrogen. The growth analysis of dry matter showed an early depressive effect of nitrogen deficiency and a similar, but less important effect concerning phosphorus.

The induced nutritional stresses showed an important increase in stomata resistance, caused primarily by nitrogen or potassium deficiency, accompanied by a reduction of chlorophyll concentration, which resulted from the removal of phosphorus, magnesium, and mainly nitrogen supply. Remarkable starch synthesis and storage was also revealed following nitrogen deficiency, but it was very weak after the suppression of magnesium, which may provoke a weakening of growth and development of these plants.     

Retention of xenon in quartz and Earth's missing xenon

The reactivity of xenon with terrestrial oxides was investigated by in situ synchrotron x-ray diffraction. At high temperature (T > 500 kelvin), some silicon was reduced, and the pressure stability of quartz was expanded, attesting to the substitution of some xenon for silicon. When the quartz was quenched, xenon diffused out and only a few weight percent remained trapped in samples. These results show that xenon can be covalently bonded to oxygen in quartz in the lower continental crust, providing an answer to the missing xenon problem; synthesis paths of rare gas compounds are also opened.     

Cell viability effects and antioxidant and antimicrobial activities of Tunisian date syrup (Rub El Tamer) polyphenolic extracts

The aqueous-acetone polyphenolic extract of the traditionally derived date syrup, known as "Rub El Tamer", was analyzed using RP-HPLC-DAD and ESI-MS. The phenolic content of the extract was 394.53 ± 1.13 mg per 100 g of syrup with caffeoylsinapylquinic acid as the most abundant compound (72.23%). The extract exhibited strong antioxidant activities as evaluated using the ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)), DPPH (2,2-diphenyl-1-picrylhydrazyl) and FRAP (ferric reducing antioxidant power) methods. The extract antimicrobial potential against a range of microorganism strains showed that Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus cereus were the most sensitive bacteria with MBC in the range of 0.5-0.05 mg/mL. Furthermore, in the presence of the syrup extract (8.18-131 μg/mL), the Human SH-SY5Y neuroblastoma and the 3T3 fibroblast cell lines showed dissimilar reduction of viability suggesting a higher cytotoxic effect against tumorigenic cells. Our results provide new insights into date syrup characterization which should stimulate further studies of this hot desert resource.     

Novel polyphenol molecule isolated from licorice root (Glycrrhiza glabra) induces apoptosis, G2/M cell cycle arrest, and Bcl-2 phosphorylation in tumor cell lines

Herbal therapies are commonly used by patients with cancer, despite little understanding about biologically active chemical derivatives. We recently demonstrated that the herbal combination PC-SPES, which contains licorice root, had anti-prostate cancer activity attributable to estrogen(s) that produced a chemical castration. A recent study also demonstrated that licorice root alone decreased circulating testosterone in men. Other studies demonstrated antitumor activity of PC-SPES in vitro associated with decreased expression of the anti-apoptotic protein Bcl-2 and in patients independent of chemical castration, suggesting that other mechanisms of antitumor activity exist separate from chemical castration. In the present study, we assessed licorice root extract for effects on Bcl-2 to identify novel cytotoxic derivatives. Licorice root extract induced Bcl-2 phosphorylation as demonstrated by immunoblot and G2/M cell cycle arrest, similarly to clinically used antimicrotubule agents such as paclitaxel. Bioassay-directed fractionations resulted in a biologically active fraction for Bcl-2 phosphorylation. HPLC separation followed by mass spectrometry and NMR identified 6 compounds. Only one molecule was responsible for Bcl-2 phosphorylation; it was identified as 1-(2,4-dihydroxyphenyl)-3-hydroxy-3-(4'-hydroxyphenyl) 1-propanone (beta-hydroxy-DHP). The effect on Bcl-2 was structure specific, because alpha-hydroxy-DHP, 1-(2,4-dihydroxyphenyl)-2-hydroxy-3-(4'-hydroxyphenyl) 1-propanone, in contrast to beta-hydroxy-DHP, was not capable of Bcl-2 phosphorylation. Pure beta-hydroxy-DHP induced Bcl-2 phosphorylation in breast and prostate tumor cells, G2/M cell cycle arrest, apoptosis demonstrated by Annexin V and TUNEL assay, decreased cell viability demonstrated by a tetrazolium (MTT) assay, and altered microtubule structure. Therefore, these data demonstrate that licorice root contains beta-hydroxy-DHP, which induced Bcl-2 phosphorylation, apoptosis, and G2/M cell cycle arrest, in breast and prostate tumor cells, similarly to the action of more complex (MW >800) antimicrotubule agents used clinically.     

Differences between wheat cultivars in acquisition and utilization of phosphorus

In an attempt to evaluate whether breeding and selection for high yielding capacity changed the P requirement of modem wheat cultivars. the response of two wheat cultivars to different levels of P supply was investigated. A traditional cultivar (‘Peragis’) and a modern spring-wheat cultivar (‘Cosir’) were cultivated in a C-loess low in available P and high in CaCO₃ in 120 cm high PVC tubes. In addition and for comparison, nutrient solution experiments were also conducted. Shoot growth, root growth. P uptake. P translocation and P distribution within the shoot at different developmental stages were compared. The grain yield of the modern cultivar ‘Cosir’ was higher at limiting and non-limiting P supply and. therefore, this cultivar can be considered as more P-efficient than the traditional cultivar. Grain yield reduction at low P supply was mainly due to an inhibition of tillering and thus lower number of ears per plant, whereas the number of grains per ear was hardly affected. Reduced tillering at low P supply could not be related to P concentrations in the shoot meristematic tissues which were generally much higher than in other plant tissues and kept at an elevated level even at limiting P supply. Root branching (1^st order laterals) was reduced at limiting P supply in ‘Cosir’ but not in ‘Peragis’ which, generally, had lower numbers of laterals at the beginning of tillering. From the results it can be concluded that the main factors contributing to the higher P efficiency of the modern cultivar ‘Cosir’ are (i) efficient use of assimilates for root-growth characteristics which enhance P acquisition: enhanced root branching and thus smaller mean root diameter and longer root hairs, (ii) an efficient P uptake system, (iii) efficient remobilization of P from vegetative plant organs to the grains, and most importantly (iv) lower P requirement for grain yield formation because of lower ear number per plant but higher grain number per ear.     

Prediction of hydraulic conductivity and sorptivity in soils at steady-state infiltration

The purpose of this study was (1) to find a matching factor (u) between infiltration rate and hydraulic conductivity during steady-state infiltration, and (2) to propose equations based on infiltration and soil moisture-retention functions for prediction of the hydraulic conductivity K(θ) within the rapidly (non-capillary) drainable pores (RDP) and capillary-matrix pores of soils. The K(θ) of capillary pores was divided into K(θ)_SDP, K(θ)_WHP and K(θ)_FCP within slowly drainable pores (SDP), water-holding pores (WHP) and fine capillary pores (FCP), respectively. Five soil profiles of calcareous sandy loam, alluvial saline and non-saline clay, located at the Nile Delta, were used to apply the proposed equations. The highest and the lowest values of K(θ)_RDP were observed in calcareous and saline clay soil profiles, respectively. Values of K(θ)_RDP remained higher than those for capillary pores in the studied soils. The predicted values of K(θ) in capillary and non-capillary pores classes were in the expected range for unsaturated hydraulic conductivity. Water sorptivity (S) was determined at initial unsaturated soil water conditions and calculated at steady-state infiltration (S _w) using a derived equation. There was a decrease in S with an increase in soil water content; i.e. at steady-state infiltration, S decreased by 35–40% in calcareous soils and by 45–60% in alluvial clay soils. The parameter values of u and S _w tended to be uniform in calcareous soils, but nonuniform in saline and non-saline clay soils.     

Optimized Extraction,Preliminary Characterization and Evaluation of the in Vitro Anticancer Activity of Phlorotannin-Rich Fraction from the Brown Seaweed,Cystoseira sedoides

Brown seaweeds produce useful bioactive substances with high cosmetic and pharmacological values due to the presence of antioxidant derivatives, mainly phlorotannins (PHT), which are of particular interest. A central composite rotatable design (CCRD) with three variables (extraction time, dry material-to-solvent ratio and ethanol concentration) and two responses was performed to optimize the microwave-assisted extraction (MAE) of phlorotannins from Cystoseira sedoides using response surface methodology (RSM). Results showed that the independent variables significantly affected both phlorotannin content and the scavenging capacity. The optimum operating conditions were extraction time, 101.74 sec; dry material-to-solvent ratio, 1:10 g/mL; and ethanol extraction, 50%. Under these conditions, the predicted values of PHT content and radical scavenging activity-IC50 were close to the observed values and were 383.887 µg PGE/g Dm and 18.353 µg/mL, respectively. Characterization of the phlorotannin-rich fraction was conducted by high-performance liquid chromatography and Fourier transform infrared spectroscopy. The evaluation of the anticancer activity against MCF-7 breast cancer cell line showed a potent activity to trigger apoptotic death in more than a half of the MCF-7 cells, with an estimated IC50 value of 78 μg/mL. In addition, this fraction induced a notable growth regression effect on 3D spheroids model in a concentration-dependent manner, with a growth rate of about 1.17, at 200 µg/mL.

Abbreviations: CCRD: Central composite rotatable design; Dm: Dry material; DMBA: 2,4-dimethoxy benzaldehyde; DMSO: Dimethyl sulfoxide; DPPH: 1,1-diphenyl-2-picrylhydrazyl; EDTA: Ethylene diamine tetraacetic acid; FBS: Fetal bovine serum; FT-IR: Fourier transform infrared; HPLC: High-performance liquid chromatography; IC₅₀: Half-maximal inhibitory concentration; MAE: Microwave-assisted extraction; Min: minutes; PBS: Phosphate buffered saline; PGE: Phloroglucinol equivalent; PHT: Phlorotannins; PHT-SED: phlorotannins derived from Cystoseira sedoides; PI: Propidium iodide; RSA: Radical scavenging activity; RSM: Response surface methodology; Sec: seconds     

Evaluation of in vitro and in vivo inhibitory effects of fusidic acid on Babesia and Theileria parasites

Fusidic acid known to has antibacterial, antifungal, and antimalarial activities. Fusidic acid blocks translation elongation factor G gene in Plasmodium falciparum. In the present study, the inhibitory effects of fusidic acid on the in vitro growth of bovine and equine Babesia parasites were evaluated. The inhibitory effect of fusidic acid on the in vivo growth of Babesia microti was also assessed. The in vitro growth of four Babesia species that were tested was significantly inhibited (P < 0.05) by micromolar concentrations of fusidic acid (IC₅₀ values = 144.8, 17.3, 33.3, and 56.25 μM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Combinations of fusidic acid with diminazene aceturate synergistically potentiated its inhibitory effects in vitro on B. bovis and B. caballi. In B. microti-infected mice, fusidic acid caused significant (P < 0.05) inhibition of the growth of B. microti at the dose of 500 mg/kg BW relative to control group. These results indicate that fusidic acid might be incorporated in treatment of babesiosis.