Analysis of Antioxidant and Antiviral Effects of Olive (Olea europaea L.) Leaf Extracts and Pure Compound Using Cancer Cell Model.

The present study aims to assess the antioxidant and antiviral effectiveness of leaf extracts obtained from Olea europaea L. var. sativa and Olea europaea L. var. sylvestris. The total antioxidant activity was determined via both an ammonium phosphomolybdate assay and a nitric oxide radical inhibition assay. Both extracts showed reducing abilities in an in vitro system and in human HeLa cells. Indeed, after oxidative stress induction, we found that exposition to olive leaf extracts protects human HeLa cells from lipid peroxidation and increases the concentration of enzyme antioxidants such as catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase. Additionally, OESA treatment affects viral DNA accumulation more than OESY, probably due to the exclusive oleuropein content. In fact, subtoxic concentrations of oleuropein inhibit HSV-1 replication, stimulating the phosphorylation of PKR, c-FOS, and c-JUN proteins. These results provide new knowledge about the potential health benefits and mechanisms of action of oleuropein and oleuropein-rich extracts.

Characterization of nettle leaves (Urtica dioica) as a novel source of protease for clotting dromedary milk by non-destructive methods.

This study investigates the valorization of the nettle leaves (Urtica dioica) as a novel source of a protease for clotting dromedary milk. The aim of this work is to study the effect of extracting pH on the enzymatic activity of nettle leaves extracts. The extraction was achieved in phosphate citrate buffer at different pH values (from 3 to 6.5) and the obtained extracts were used to coagulate dromedary milk. The characterization of the obtained extracts was carried out using non-destructive methods namely FT-MIR, fluorescence spectroscopy and turbiscan instrument. The extract prepared at pH = 4 had the highest proteolytic activity. The fluorescence and turbiscan measurements revealed a substantial effect of the pH value on chlorophyll residues extraction and stability, respectively. At an acidic environment (pH range of 3 - 4), the enzymatic extracts were unstable (with turbiscan stability index (TSI) values ~ 20), while at a nearly neutral pH value (pH range of 5 - 6.5), they were found to be more stable as indicated by the low TSI values ~ 1. The maximum milk-clotting activity (MCA) (0.021 U/mL) was obtained for the extracts prepared at pH = 4.

In Vitro Anti-Epstein Barr Virus Activity of Olea europaea L. Leaf Extracts.

Olea europaea L. var. sativa (OESA) preparations are widely used in traditional medicine in the Mediterranean region to prevent and treat different diseases. In this research, olive extracts derived from the leaves of the OESA tree have been screened for antioxidant activity by two methods: the DPPH free radical scavenging assay (DPPH) and the Ferric reducing antioxidant power (FRAP) assay. The DPPH assay showed that OESA possesses a stronger antioxidant activity (84%) at 1 mg/mL while the FRAP method showed a strong metal ion chelating activity (90%) at 1 mg/mL. The low IC50 values, obtained by two different methods, implies that OESA has a noticeable effect on scavenging free radicals comparable to standards. During EBV infection, the free radicals increased triggering lipid oxidation. Therefore, the monitoring of the secondary lipid peroxidation products was done by measuring malonaldehyde (MDA) and conjugated dienes (DC). The simultaneous treatment of Raji cells with OESA and TPA, as an inductorof the lytic cycle, generated a significant decrease in MDA levels and DC (p < 0.05). Besides, Raji cells simultaneously exposed to TPA and OESA exhibited a percentage of EBV-positive fluorescence cells lower than TPA treated cells (**** p < 0.0001). This suggests that OESA treatment has a protective effect against EBV lytic cycle induction.

Development and characterization of chitosan films carrying Artemisia campestris antioxidants for potential use as active food packaging materials.

Active food packaging films based on chitosan and enriched with Artemisia campestris hydroalcoholic extract (ACHE), aqueous extract (ACAE) and essential oil (ACEO) were developed. The effects of incorporating A. campestris were investigated on the physical, mechanical, thermal and antioxidant characteristics of the films. The structural properties of the films were evaluated using Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD) and scanning electron microscopy (SEM). The results showed that adding ACHE and ACEO improved the water resistance of chitosan films. The FTIR spectroscopy analysis revealed covalent interaction and hydrogen bonding between chitosan and ACHE. The XRD and SEM analyses indicated that interactions occurred between the film matrix and A. campestris active compounds, which could be reflected by the physical and mechanical properties of composite films. Incorporating ACHE and ACAE in the chitosan matrix decreased the tensile strength. The film extensibility was reduced when ACHE and ACEO were added. All films exhibited great thermal stability as the degradation occurred above 300 °C. The addition of A. campestris active compounds, particularly extracts, to chitosan films notably increased the antioxidant and UV-Vis barrier properties. Chitosan films enriched with the A. campestris antioxidant compounds could be applied as food packaging alternatives.

Effect of extraction procedures on the chemical structure, antitumor and anticoagulant properties of ulvan from Ulva lactuca of Tunisia coast.

The effect of extraction procedures on chemical composition, structural, antitumor and anticoagulant properties of the sulphated polysaccharide 'ulvan' from the green seaweed Ulva lactuca were investigated. The structural features of ulvans were carried out by FTIR and by one- and two- dimensional 1H and 13C NMR spectroscopic. The ulvans were mainly composed of rhamnose, xylose, and uronic acid. Chemical and spectroscopic analyses demonstrated that ulvans were constituted of (1→4)-ß-glucuronic acid, (1→3,4)-α-L-rhamnose-3-sulphate and (1→4)-α-xylose. The extraction procedures effect were observed in chemical structure, Mw and biological activities. Cytotoxic activity of enzymatic-chemical extract on cervical cancer cells (HeLa) (IC50 = 1000 µg/mL) was higher than on normal peripheral blood lymphocytes cells (PBL). Acid extracts promoted to reduce HeLa cells and to grow PBL cells. At high concentrations, acid extracts showed the highest APTT and TT clotting time. Antitumoral and anticoagulant activities of ulvans from Ulva lactuca promote their use as effective therapeutic agent.

Effect of extraction methods on the physicochemical, structural, functional, and antioxidant properties of the dietary fiber concentrates from male date palm flowers.

The current study evaluates the effect of both aqueous and alkaline extraction methods on the physicochemical, structural, functional, and antioxidant properties of dietary fibers (DF) from male date palm flowers (MDPF). The DF extracted by the alkali treatment (ADF) display a higher DF content as well as lower protein and lipid content than the DF resulting from watery extraction (WDF). The scanning electron microscopy, the Fourier-transform infrared spectroscopy, and the X-ray diffraction show that the alkaline treatment contributes to a slight modification of the structural characteristics of ADF leading to better purity and functional properties. The oil holding capacity and the antioxidant activity of ADF have improved compared to the WDF, which makes this concentrate a promoting functional ingredient and a natural antioxidant. Therefore, these findings confirm the wealth of both concentrates in DF, especially ADF, as well as their potential of functional and antioxidant properties, which emphasize their suitability to be used in food applications as functional ingredients. PRACTICAL APPLICATIONS: Nowadays, studies on the extraction of DF from vegetable waste are of considerable interest in the face of the rising global demand for dietetic food. The male date palm flowers (MDPF), an agricultural waste generated during the period of pollination, are valuable natural sources of DF and antioxidants. The insoluble DF of MDPF could be essential in a balanced diet as they could fight against obesity by creating a feeling of satiety. The presence of antioxidants, which are known for preventing or slowing cell damage caused by free radicals, could contribute to the improvement of the antioxidant properties of the formulated food. The functional properties of DF concentrates from MDPF are able to solve the technical problems of agri-food industries when used as food ingredients. The use of DF from MDPF, consequently, contributes to the minimization of waste and provides value addition to the by-product considered as waste in agricultural processing.

Physico-Chemical, antioxidant activities, textural, and sensory properties of yoghurt fortified with different states and rates of pomegranate seeds (Punica granatum L.).

This study examines the effects of incorporating fresh, frozen, osmodehydrated, and dried pomegranate seeds, at different concentrations (5, 10, and 20%) on the physicochemical, antioxidant, textural, and sensory properties of yoghurt. Antioxidant activity, pH, acidity, syneresis, and color of yoghurt were also evaluated during the storage (28 days) at 4°C. The principal component analysis (PCA) was performed to assess the correlations between different yoghurt formulation and their sensory attributes. Data showed that the addition of pomegranate seeds reduced pH and modified the chromatics coordinate (L*, a*, b*, C*, h°) and firmness of the yoghurt samples. In addition, acidity, °Brix, and syneresis increased compared to the control. Thanks to their high anthocyanin content, pomegranate seeds considerably improved the antioxidant activity essentially for yoghurt enriched with frozen seeds. In addition, the supplementation of 20% of frozen seeds into yoghurt was the most appreciated by panelists and improved the sensory properties in comparison to other formulation. According to PCA, taste and appearance were the main criteria for the overall acceptability of yoghurt. After 28 days of storage, the reddish color was reinforced; however, the antioxidant activity of yoghurts was reduced. Overall, it can be concluded that pomegranate seeds can be used as a natural ingredient to develop a novel yoghurt with high nutritional properties.

Quality Characteristics and Functional and Antioxidant Capacities of Algae-Fortified Fish Burgers Prepared from Common Barbel (Barbus barbus).

INTRODUCTION: Algae have been used as natural ingredients to produce new canned fish burgers prepared from minced flesh of common barbel. In this research, the impact of the addition of Cystoseira compressa and Jania adhaerens at concentrations of 0.5, 1, and 1.5% w/v on the texture and sensory characteristics of fish burgers were investigated. RESULTS: Compared to controls, fish burgers containing 1% algae had better texture and sensory properties (P < 0.05). Also, these burger formulations had higher water and oil holding capacities as well as swelling ability, due to the important polysaccharides and dietary fibers contents of algae. In addition, algae-supplemented burgers were characterized as having low L⁎, a⁎, and b⁎ values, which made the color appear to be paler. Thanks to their high richness in pigments (chlorophylls and carotenoids) and polysaccharides, algae considerably enhance the antioxidant activities of the new ready-to-eat fish burgers. So, Cystoseira compressa and Jania adhaerens could be used as nutritious additives to produce new fish-based products.

Physicochemical and rheological changes of acidified camel milk added with commercial low methoxyl-pectin.

The influence of the addition of low methoxyl amidated pectin (LMA) on acid milk gels from whole camel milk (WCM) on physicochemical and rheological proprieties were studied. The zeta potential, particle size, viscosity, dynamic oscillatory rheology and isothermal titration calorimetry were monitored. The presence of LM-pectin in milk had an impact on the average size of the casein micelles and a large and dominant influence on rheological behavior during acidification. Zeta potential and viscosity of gels with 0.5% pectin were not affected. However, milk gels containing 1%, 1.5% and 2% of LM-pectin showed highest values of particle size at pH4. This modification of the structure of the casein micelles induces a significant improvement (p<0.05) on its acid gelation behavior. Therefore, the addition of pectin enhanced the rheological proprieties. Higher pectin concentration led to a strong gel with higher G' values. This result could be attributed to the formation of complexes and the mechanical spectra prove the hypothesis that pectin forms strands with caseins micelles. Isothermal titration calorimetry results showed that pectin concentration had a marked influence on the gels structure and that this polysaccharide stabilizes caseins micelles in acidified camel milk gel due to electrostatic interaction and steric repulsion.

Toward the enhancement of sensory profile of sausage "Merguez" with chickpea protein concentrate.

The influence of different Chickpea Protein Concentrates (CPC) addition at different levels (CPC: 1.5, 2.5 and 5% (w/w)) on physico-chemical, textural and hedonic properties of formulated and cooked sausage "Merguez" were investigated. The obtained results exhibited that CPC addition increased protein content of "Merguez" sausage and improved the process yield. Significant differences were noticed on the cooking loss and textural properties for "Merguez" sausages formulated with 5% of CPC compared to the control (p < 0.05). Hedonic analysis showed that protein addition had no significant effect on sausages taste. However, sausage texture and global acceptability were markedly improved (p < 0.05). These results suggest that CPC could be an important alternative source of protein additive for the improvement of the physico-chemical, textural and sensorial properties of meat sausages.

Ultrafiltration and thermal processing effects on Maillard reaction products and biological properties of date palm sap syrups (Phoenix dactylifera L.).

The effect of ultrafiltration process and temperature concentration on MRPs content and antioxidant, antimicrobial and cytotoxic properties of date palm sap syrups were investigated. MRPs were analyzed by HPLC. Antioxidant activity was evaluated by reducing power and DPPH free radical and H2O2 scavenging activities. Antimicrobial activity was evaluated by the agar disk diffusion method. In vitro cytotoxic activity was examined by cell proliferation assay. Date sap syrups displayed strong antioxidant activities which are correlated 5HMF and 2F contents. In addition, concentration at 100 °C, unlike ultrafiltration process, enhanced significantly the antioxidant activities sap syrups and total phenolic contents. The antimicrobial activities showed marked activity against S. enterica, P. aeruginosa, S. aureus, L. monocytogenes with an inhibition zone of 21, 34, 27 and 34 mm respectively. Cytotoxicity assays showed that sap syrups can inhibit the proliferation of HeLa cell lines at high concentration.

Influence of the ripening stage and the lyophilization of wild cardoon flowers on their chemical composition, enzymatic activities of extracts and technological properties of cheese curds.

The selection of an appropriate Cynara cardunculus flowers batch is a preliminary step to produce rennet with better clotting properties. For this reason, we proposed to study the influence of the ripening stage and the lyophilization of cardoon flowers on their chemical composition, enzymatic activities of extracts, and technological properties of cheese curds. Results of flowers composition have shown that lyophilized flowers harvested at the middle of ripening stage (A) could be employed to produce mainly proteins or milk-clotting proteases. To confirm this, enzymatic activities of extracts and technological properties of curds were assessed. The experimental findings revealed that flowers lyophilization seems to be an efficient way to produce rennet with better clotting properties, leading to higher yield, moisture, and texture parameters of curd. These findings allowed us to select lyophilized flowers (A) for further cheese making process.

Storage stability of traditional Tunisian butter enriched with antioxidant extract from tomato processing by-products.

Traditional Tunisian butter (TTB) is one of the most appreciated dairy products in Tunisia. Herein, the storage stability of TTB enriched with antioxidants from tomato processing by-products (TPB) was evaluated during 60days of storage at 4°C. TPB extract contains significant amounts of lycopene and phenolics. TTB enriched with 400mg of TPB extract/kg of TTB revealed the lowest peroxide values at all the determination intervals. Adding 400mg of TPB extract/kg of TTB did not exhibit any undesired effect on lactic bacteria which are necessary for development of aroma and chemical properties of TTB. However, raw TTB and highly enriched TTB (800mg of TPB extract/kg of TTB) displayed higher lipid peroxidation. The detrimental effect of high antioxidant amounts on TTB stability could be due to a possible pro-oxidant character. Thus, appropriate supplementation of TPB extract could be used in TTB as a protective agent against lipid peroxidation to extend its shelf-life up to two months.

Characterization of pectins extracted from pomegranate peel and their gelling properties.

The composition of pomegranate peel, the main by-product during pomegranate processing, and some of the characteristics of the water-soluble pectins were investigated. Four tunisian pomegranate peels were subjected to hot aqueous extractions (86°C, 80min, 20mM nitric acid). Pomegranate peels yielded between 6.8% and 10.1% pectins. The extracted pectins were low methylated and were characterized by the predominance of homogalacturonan regions. Principal component analysis applied on FT-IR spectral data in the region between 4000 and 650cm(-1) differentiated the samples according to their degree of methylation. At pH 3, in the presence of 0.7% pectin, all solutions showed a rapid gel formation with G'>G″. With decreasing temperature from 90°C to 10°C, G' increased to reach a plateau at 10°C. The variation in the pectin gel formation between varieties was attributed to difference in pectin characteristics particularly the hydrodynamic volume and the neutral sugar content.

Yield and composition of pectin extracted from Tunisian pomegranate peel.

A central composite design was employed to determine the influence of extraction conditions on production yield and chemical composition of pectin from pomegranate peels. Response surface methodology (RSM) was used to quantify the integral effect of the three processing parameters (extraction duration, temperature and pH) on yield. A second-order polynomial model was developed for predicting the yield of pomegranate peels pectin based on the composite design. Yields ranged from 6.4 to 11.0±0.2%. Optimal temperature, duration and pH value of the extraction were 86°C, 80min and 1.7, respectively. The uronic acid and the total neutral sugar content of the extracted pectins ranged from 377 to 755mg/g and from 161 to 326mg/g, respectively. Moreover, the degree of methylation varied with the extraction conditions and the extracted pectins were low methylated. On high pressure size exclusion chromatography (HPSEC), the elution pattern of the acid-extracted pectins showed that severe conditions were associated with lower hydrodynamic volume.

Effect of ultrafiltration process on physico-chemical, rheological, microstructure and thermal properties of syrups from male and female date palm saps.

This study investigates the effect of the ultrafiltration process on physicochemical, rheological, microstructure and thermal properties of syrups from male and female date palm sap. All the studied syrups switched from pseudoplastic rheological behaviour (n=0.783) to Newtonian behaviour (n∼1) from 10 to 50 °C respectively and present similar thermal profiles. Results revealed that the ultrafiltration process significantly affects the rheological behaviour of the male and female syrups. These differences on rheological properties are attributed to the variation of chemical composition between sap and sap permeate syrups. Furthermore, the effect of temperature on viscosity of the syrups was investigated during heating and cooling processes at the same shear rate (50s(-1)). This study provides idea of the stability of the syrup by evaluating the area between heating and cooling curves. Actually, the syrup prepared from male sap permeate is the most stable between the four studied syrups.

Phenolic profile, antibacterial and cytotoxic properties of second grade date extract from Tunisian cultivars (Phoenix dactylifera L.).

The present study aimed to investigate the phenolic profile of second grade date extracts and evaluate their antimicrobial and cytotoxic activities with regard to some pathogenic microorganisms. Phenolic content was analyzed by HPLC. Antimicrobial activity was evaluated by the agar disk diffusion method, and in vitro cytotoxic activity was examined by cell proliferation assay. The results revealed that second grade dates presented three benzoic acids, five cinnamic acids and two flavonoids, with the predominance of ρ-coumaric acid (1998.80µg/100g). The antimicrobial activities showed that the date extracts were active against Gram (+) and Gram (-) bacteria, showing marked activity against Escherichia coli with an inhibition zone of 25mm. Cytotoxicity assays showed that the date extracts were able to inhibit the proliferation of HeLa cell lines. The results confirmed that the date extracts were rich in biologically active compounds that are highly valued in the functional food and nutraceutical industries.

Antioxidant, antibacterial and in vivo dermal wound healing effects of Opuntia flower extracts.

Opuntia ficus-indica flowers are used for various medicinal purposes. The aims of the present investigation were to evaluate biological properties of O. ficus-indica flowers extracts and to investigate its antioxidant and antibacterial activities and its ability to enhance wound healing. The wound healing activity of the mucilaginous and methanol extracts of O. ficus-indica flowers were assessed using excision wound model in rats. After thirteen days of treatment by both extracts, a beneficial effect on cutaneous repair was observed as assessed by the acceleration of wound contraction and remodeling phases. Histopathological studies of the granulation tissue indicated that the derma is properly arranged with the Opuntia flowers extract, compared with the control group. The mucilage extract was more effective than the methanol extract, but both showed significant results compared with the control. Such investigation was supported by the efficiency of the methanolic and mucilage extract as antimicrobial and antioxidant. Indeed, the extracts showed a potential antioxidant activity determined by different test systems, namely DPPH radicals scavenging activity, trolox equivalent antioxidant capacity, reducing power, ß-carotene bleaching assay and metal chelating activity and exhibited significant antibacterial activity against almost all tested bacteria.

Phenolic Profiles, Phytchemicals and Mineral Content of Decoction and Infusion of Opuntia ficus-indica Flowers.

Opuntia flowers are a natural source of biologically active compounds and they have been used as medicinal plant for a long time. Despite the various uses reported for the decoction and infusion of these flowers, their characterization has been discarded. In this study, the decoction and infusion prepared from Opuntia ficus-indica were analyzed with respect to their content in minerals and phytochemicals in order to evaluate its nutritional characteristics. The obtained data proved that these preparations are a rich source of minerals mainly K and Ca. Moreover, the phytochemical analysis revealed that they have important polyphenols, flavonoids and tannins contents with the infusion that presented the highest polyphenol levels. LC-MS analyses of decoction and infusion allowed the characterization of 20 phenolic compounds. It is mainly identified by the presence of flavonols glycosides.

Chemical composition and antibacterial activity of Opuntia ficus-indica f. inermis (cactus pear) flowers.

Opuntia ficus-indica f. inermis (cactus pear) flowers have wide application in folk medicine. However, there are few reports focusing on their biological activity and were no reports on their chemical composition. The nutrient composition and hexane extracts of Opuntia flowers at 4 flowering stages and their antibacterial and antifungal activities were investigated. The chemical composition showed considerable amounts of fiber, protein, and minerals. Potassium (K) was the predominant mineral followed by calcium (Ca), magnesium (Mg), sodium (Na), iron (Fe), and zinc (Zn). The main compounds in the various hexane extracts were 9.12-octadecadienoic acid (29-44%) and hexadecanoic acid (8.6-32%). The antibacterial activity tests showed that O. inermis hexane extracts have high effectiveness against Escherichia coli and Staphylococcus aureus, making this botanical source a potential contender as a food preservative or food control additive.