Comparative study of the effect of oleuropein and hydroxytyrosol rich extracts on the reproductive toxicity induced by bisphenol A in male rats: biochemical, histopathological, and molecular analyses.

Bisphenol A, or BPA, goes into the composition of a large number of products including sunglasses, infant's feeding bottles, receipts, or food packaging. Nowadays, there is a growing evidence that BPA may be at the origin of several physiological malignancies. Oleuropein and hydroxytyrosol extracted from olive leaves are highly investigated for numerous health benefits. The present work investigates the potential protective proprieties of olive leaf extracts against BPA-induced testicular damage in Wistar rats. Thirty-two animals were randomly divided into 4 groups: control, BPA-treated (10 mg/kg), BPA and oleuropein rich extract (16 mg/kg) treatment, and the last group treated with BPA and hydroxytyrosol rich extract (16 mg/kg). Biochemical parameters and histological and molecular analyses were evaluated. Our data demonstrated that BPA treatment caused significant alteration in biochemical parameters, disorganization of germinal epithelium, an up-regulation of p53 and Bax, and a reduction of Bcl-2 protein levels. The ingestion of oleuropein- and hydroxytyrosol-rich extracts attenuated BPA-induced biochemical and histological changes. In fact, olive leaf extracts enhanced the enzymatic antioxidant system and the level of Bcl-2, and reduced the expression of p53 and Bax. Fairly, our findings propose that olive leaf extracts may compete with BPA-induced reprotoxicity in vivo.

Olive Leaves Extract and Oleuropein Improve Insulin Sensitivity in 3T3-L1 Cells and in High-Fat Diet-Treated Rats via PI3K/AkT Signaling Pathway.

Olive leaves extracts are known to exert potential pharmacological activities especially, antidiabetic and antiobesity. This study explores the anti-insulin resistant effect of olive leaves extracts and oleuropein in 3 T3-L1 cells and in high-fat diet fed rats. Our results showed that ethanol extract (EE) suppressed significantly (P < 0.01) triacylglycerol accumulation. In preadipocytes cells, EE 1/100 decreased cell viability and induced apoptosis. Real-time PCR analysis showed that EE reduced the mRNA levels of adipogenesis (CEBP-α, PPARγ, SREBP-1c, and FAS) and proinflammatory (TNF-α and IL-6) genes. Moreover, the cotreatment of EE 1/1000 or oleuropein with insulin increased considerably the expression of p-IRS, p85-pI3K, and p-AKT. In vivo model, the oral administration of oleuropein at 50 mg/kg in rats fed with high fat diet for 8 weeks reduced inflammation in liver and adipose tissues (WAT), improved glucose intolerance, and decreased hyperinsulinemia. Furthermore, the immunohistochemistry revealed that the expression level of p-Akt, IRS1, and Glut-4 were significantly enhanced in liver and WAT tissues after oleuropein supplementation comparing with that in HFD group. Additionally, the expression of IRS1 was markedly ameliorated in pancreas. Our obtained results can be adopted as an approach to used olive leaves as complement to prevent insulin-resistance disease.

Optimizing the Extraction Conditions of Hydroxytyrosol from Olive Leaves Using a Modified Spherical Activated Carbon: A New Experimental Design.

The purification of hydroxytyrosol from olive leaves extract by modified activated carbon was studied experimentally in a batch system and a column by adsorption and desorption processes. The extraction yield reached 90% of hydroxytyrosol, which is the major compound found in the extract. Despite the abundance of research on extracts of hydroxytyrosol from olive leaves, it seems that the applied methods can be further improved. In this study, several approaches were applied to optimize the extraction conditions of this molecule. Hence, the response surface method and the Box-Behnken design (BBD) were used to evaluate the effect of the temperature, time, and adsorbent dose on the hydroxytyrosol recovery. Moreover, adsorption isotherm, kinetics, and thermodynamic studies were also performed to clarify the nature of the process. The main finding was the obtainment of a maximum adsorption yield of 97.5% at an adsorbent/adsorbate ratio of 1 : 20, after a 6 h cycle and at a temperature of 30°C. Furthermore, adsorption process seemed to fit best with Freundlich model. In addition, the thermodynamic study describes a spontaneous and endothermic process. Desorption assay using ethanol helped to recover 73% of hydroxytyrosol. Furthermore, the HPLC analysis of fractions after column adsorption showed a simple peak of hydroxytyrosol with purity higher than 97% and a flavonoids-rich fraction. These findings would indicate that this separation method for the recovery of phenolic compounds with high antioxidant activity can be a very promising one.

Protective effect of olive leaves phenolic compounds against neurodegenerative disorders: Promising alternative for Alzheimer and Parkinson diseases modulation.

The main objective of this work was to review literature on compounds extracted from olive tree leaves, such as simple phenols (hydroxytyrosol) and flavonoids (Apigenin, apigenin-7-O-glucoside, luteolin.) and their diverse pharmacological activities as antioxidant, antimicrobial, anti-viral, anti-obesity, anti-inflammatory and neuroprotective properties. In addition, the study discussed the key mechanisms underlying their neuroprotective effects. This study adopted an approach of collecting data through the databases provided by ScienceDirect, SCOPUS, MEDLINE, PubMed and Google Scholar. This review revealed that there was an agreement on the great impact of olive tree leaves phenolic compounds on many metabolic syndromes as well as on the most prevalent neurodegenerative diseases such as Alzheimer and Parkinson. These findings would be of great importance for the use of olive tree leaves extracts as a food supplement and/or a source of drugs for many diseases. In addition, this review would of great help to beginning researchers in the field since it would offer them a general overview of the studies undertaken in the last two decades on the topic.

Investigation of halotolerant marine Staphylococcus sp. CO100, as a promising hydrocarbon-degrading and biosurfactant-producing bacterium, under saline conditions.

A halotolerant strain CO100 of Staphylococcus sp. was isolated from contaminated sediments taken from the fishing harbour of Sfax, Tunisia, as an efficient hydrocarbonoclastic candidate. Strain CO100 exhibited a high capacity to break down almost 72% of the aliphatic hydrocarbons contained in crude oil (1%, v/v), used as the sole carbon and energy source, after 20 days of culture, at 100 g/l NaCl, 37 °C and 180 rpm. The isolate CO100 displayed also its ability to grow on phenanthrene, fluoranthene and pyrene (100 mg/l), at 100 g/l NaCl. Moreover, the isolate CO100 showed a notable aptitude to synthesize an efficient tensioactive agent namely BS-CO100, on low-value substrates including residual frying oil and expired milk powder, thus reducing the high cost of biosurfactant production. The ESI/MS analysis designated that BS-CO100 belonged to lipopeptide class, in particular lichenysin and iturine members. Critical micelle concentrations of BS-CO100 were varying between 65 and 750 mg/l, depending on of the purity of the biosurfactant and the used carbon sources. BS-CO100 showed a high steadiness against a wide spectrum of pH (4.3-12), temperature (4-121 °C) and salinity (0-300 g/l NaCl), supporting its powerful tensioactive properties under various environmental conditions. Likewise, BS-CO100 exhibited no cytotoxic effect toward human HEK293 cells, at concentrations within 125 and 1000 µg/ml. Furthermore, the biosurfactant BS-CO100 exhibited remarkable anti-adhesive and anti-biofilm activities, being able to avoid and disrupt the biofilm formation by certain pathogenic microorganisms. In addition, BS-CO100 was found to have more potential to remove hydrocarbons from contaminated soils, compared to some chemical surfactants. In light of these promising findings, strain CO100, as well as its biosurfactant, could be successfully used in different biotechnological applications including the bioremediation of oil-polluted areas, even under saline conditions.

Contribution of Major Polyphenols to the Antioxidant Profile and Cytotoxic Activity of Olive Leaves.

AIMS: This study was designed to investigate the phytochemical profile and the cytotoxic activities of the eco-friendly extracts of olive leaves from Chemlali cultivar. MATERIALS AND METHODS: The Phenolic composition of olive leaves extracts, the antioxidant activity and the cytotoxic effects against MCF-7 and HepG2 cells were determined. RESULTS: Olive leaves extracts showed relevant total polyphenols contents. Oleuropein was the major detected phenolic compound reaching a concentration of 16.9 mg/ml. The antioxidant potential of the studied extracts varied from 23.7 to 46.5mM Trolox equivalents as revealed by DPPH and ABTS assays. Cytotoxicity experiments showed similar trends for both HepG2 and MCF-7 cells with the infusion extract being the most active. CONCLUSION: This study denotes that olive leaves may have great potential as endless bioresource of valuable bioactive compounds which may have a wide application.

Production, characterization and biotechnological potential of lipopeptide biosurfactants from a novel marine Bacillus stratosphericus strain FLU5.

This work aimed at studying the potential of a new hydrocarbonoclastic marine bacterium, Bacillus stratosphericus FLU5, to produce an efficient surface-active agent BS-FLU5. Biosurfactant production was examined on different carbon sources; using the surface tension measurement and the oil displacement test. Strain FLU5 showed its capacity to produce biosurfactants from all tested substrates, in particular the residual frying oil, which is a cheap renewable carbon source alternative, thus minimizing the high cost of producing those surfactants. MALDI-TOF MS/MS analysis confirmed the presence of lipopeptides, which are identified as members of surfactin and pumilacidin series. The critical micelle concentration (CMC) of the purified lipopeptides produced by strain FLU5 was 50 mg/l. At this concentration, the surface tension of the water was reduced from 72 to 28 mN/m. Furthermore, the crude lipopeptides showed an interesting stability against a broad range of pH, temperature and salinity. In addition, the application of BS-FLU5 in oil recovery from hydrocarbons-contaminated soil (used motor oil) showed that it was more effective on the hydrocarbon-remobilization than some tested synthetic surfactants. Interestingly, the biosurfactant BS-FLU5 showed a negligible cytotoxic effect against the mammalian cells HEK293. These results highlight the applicability of the lipopeptides BS-FLU5 in different fields, especially in environmental remediation processes.

Oleuropein and hydroxytyrosol rich extracts from olive leaves attenuate liver injury and lipid metabolism disturbance in bisphenol A-treated rats.

In the present study, we investigated the protective effects of oleuropein- and hydroxytyrosol-rich extracts obtained from olive leaves against bisphenol A (BPA)-induced hyperlipidemia and liver injury in male rats. For this purpose, four groups of male rats (8 per group) were used: control group (Control), rats treated with BPA, rats treated with both BPA and oleuropein (OLE-BPA), and rats treated with both BPA and hydroxytyrosol (HYT-BPA). After 60 days of treatment, the results obtained using the DXA technique showed that treatment with BPA (10 mg per kg b.w.) increased the body weight and adipose tissue mass in male rats. Moreover, plasma levels of triglycerides, total cholesterol, LDL-cholesterol, AST, ALT, LDH, and TNF-α increased. The immunohistochemical analysis revealed a significant increase in the expression of COX-2 and p53 and a decrease in the expression of Bcl-2 related to liver inflammation. Oral administration of oleuropein and hydroxytyrosol-rich extracts obtained from olive leaves at 16 mg kg-1 reduced both the body weight and adipose tissue mass. These extracts were able to ameliorate liver damage and improve the elevated levels of TG and liver enzymes of BPA-treated rats possibly through enhancing CAT and SOD activities. Western blot results revealed that administration of the abovementioned extracts decreased the protein expression of NF-κB and TNF-α through the p38 signaling pathway. Overall, the findings suggest that the olive leaf extracts possess hypolipidemic and hepatoprotective effects against BPA-induced metabolic disorders through enhancing the antioxidative defense system and regulating the important signaling pathway activities.

Apigetrin inhibits adipogenesis in 3T3-L1 cells by downregulating PPARγ and CEBP-α.

BACKGROUND: Apigetrin, a flavonoid found in many plant leaves and seeds, has been known to possess antimutagenic, anti-cancer, antioxidant and anti-inflammatory properties. Here, we are investigating the effect of the apigetrin on adipocytes differentiation in 3T3-L1 adipocytes, and elucidating the mechanism of its action. METHODS: Lipids accumulation was measured by Oil Red O staining and cell cycle was analyzed by flow cytometry. The antioxidant effect of apigetrin was evaluated against hydrogen peroxide. The expression of various genes, involved in adipogenesis and inflammation, was studied by real-time PCR. RESULTS: Our results showed that apigterin treatment inhibited significantly lipid accumulation without effect on cell viability at 100 µM, and it exerted the anti-adipogenic effect during the early stages of differentiation. Flow cytometry analysis showed that apigenin-7-O-glucoside (Ap7G) inhibited cell proliferation during mitotic clonal expansion and caused cell cycle delay. Quantitative PCR analysis revealed that the mRNA levels of C/EBP-α, PPAR-γ, SREBP-1c and FAS were suppressed after apigetrin treatment at 100 µM. Moreover, the mRNA level of pro-inflammatory genes (TNF-α and IL-6) were suppressed after apigterin treatment, at high concentration preadipocyte cells. CONCLUSION: Taken together, these results indicated that apigenin-7-O-glucoside inhibits adipogenesis of 3T3-L1 preadipocytes at early stage of adipogenesis.

Effect of bacterial lipase on anaerobic co-digestion of slaughterhouse wastewater and grease in batch condition and continuous fixed-bed reactor.

BACKGROUND: This study aimed to investigate the effects of bacterial lipase on biogas production of anaerobic co-digestion of slaughterhouse wastewater (SHWW) and hydrolyzed grease (HG). A neutrophilic Staphylococcus xylosus strain exhibiting lipolytic activity was used to perform microbial hydrolysis pretreatment of poultry slaughterhouse lipid rich waste. RESULTS: Optimum proportion of hydrolyzed grease was evaluated by determining biochemical methane potential. A high biogas production was observed in batch containing a mixture of slaughterhouse composed of 75% SHWW and 25% hydrolyzed grease leading to a biogas yield of 0.6 L/g COD introduced. Fixed bed reactor (FBR) results confirmed that the proportion of 25% of hydrolyzed grease gives the optimum condition for the digester performance. Biogas production was significantly high until an organic loading rate (OLR) of 2 g COD/L. d. CONCLUSION: This study indicates that the use of biological pre-treatment and FBR for the co-digestion of SHWW and hydrolyzed grease is feasible and effective.

Assessment of Olea europaea L. fruit extracts: Phytochemical characterization and anticancer pathway investigation.

Olea europaea L. has been widely used as an advantageous rich source of bioactive compounds of high economic value leading to its use in pharmaceutical, cosmetic, and agriculture industries. Ethanolic extracts of olive fruits from three different cultivars (OFE) were studied for their phytochemical contents and were investigated for antioxidant activities and anticancer potential. Major polyphenols detected in these extracts were tyrosol, hydroxytyrosol, oleuropein, rutin, quercetin and glucoside forms of luteolin and apigenin. All these compounds have shown to significantly contribute to the antioxidant activity of OFE, which was evaluated by DPPH and ABTS assays. Proliferation of hepatic and colon cancer cells, HepG2 and Caco-2, were shown to be sensitive to OFE with IC50 less than 1.6mg/ml for all tested extracts. Moreover, flow cytometry analysis showed that OFE induced cell cycle arrest in the S-phase within both HepG2 and Caco-2 cells. This has triggered a cell death mechanism as shown by DNA fragmentation, expression of p53 and phosphorylation level of Akt and Erk proteins. Interestingly, these extracts could be further used as a potential source of natural compounds with both antioxidant and anticancer effects.

Evaluation of hypocholesterolemic effect of oleuropein in cholesterol-fed rats.

Oleuropein, which is the major compound of olive leaves, has been reported to exert several pharmacological properties, including anti-cancer, antidiabetic and anti-atherosclerotic activities. The objective of this study was to evaluate the effect of oleuropein on adiponectin level in high cholesterol diet (HCD) induced obesity in rat and the molecular mechanism underlying its activation. Our results showed that orally administered oleuropein (50 mg/kg) by gavage for 8 weeks decreased the body weight, adipose tissue mass and triglyceride and attenuated steatosis in liver. Moreover, the effect of oleuropein on adiponectin, an important hormone with fatty-acid oxidation properties, was evaluated and our data illustrated that oleuropein supplementation increased serum adiponectin concentration. The effects of oleuropein on protein expression related to lipogenic genes were investigated and our results showed that its administration significantly inhibited peroxisome proliferator-activated receptor γ (PPARγ), sterol regulatory element-binding protein-1c (SREBP-1c) and fatty-acid synthase (FAS). In addition, oleuropein stimulated the HCD-induced inhibition of AMP-activated protein kinase (AMPK) in epididymal adipose tissues. These results suggest that oleuropein exerts anti-obesity effects in HCD rats by activating AMPK and suppressing PPAR γ (Peroxisome proliferator-activated receptor γ) expression in adipose tissues. These data provide that oleuropein has important implications for preventing obesity.

Oleuropein activated AMPK and induced insulin sensitivity in C2C12 muscle cells.

AIMS: Oleuropein has been recognized as an important medicinal compound because of its various biological properties, including anti-cancer, antidiabetic and anti-atherosclerotic activities. Here, we evaluate the antioxidant activity as well as the mechanism of the hypoglycemic effects of oleuropein in C2C12 cells and we establish the mechanism underlying these effects. MAIN METHODS: To perform this study, C2C12 cells viability was analyzed via MTT assay and the antioxidant activity was investigated by ROS and TBARS assays. Also, the effect of oleuropein on AMPK and PI3 kinase signaling pathways was evaluated. KEY FINDINGS: Treatment with oleuropein was able to protect cells against H2O2 induced stress in cells. On the other hand, the molecular bases of its actions have been scarcely understood. Oleuropein significantly enhanced glucose consumption and the phosphorylation of AMPK (AMP-activated protein kinase/ACC (acetyl-CoA carboxylase)) and MAPKs (mitogen-activated protein kinases), but not PI3 kinase (Phosphatidylinositol 3-kinase)/Akt. However, the co-treatment of oleuropein and insulin improved the insulin sensitivity via insulin-dependent (PI3 kinase/Akt) and insulin independent (AMPK/ACC) pathways. These results could be confirmed from the findings of GLUT4 translocation which was strongly enhanced in the case of oleuropein. SIGNIFICANCE: Our results provide important insights for the possible mechanism of action of oleuropein as a therapeutic agent in diabetic patients.

The α-Glucosidase and α-Amylase Enzyme Inhibitory of Hydroxytyrosol and Oleuropein.

To date, numerous studies have reported on the antidiabetic properties of various plant extracts through inhibition of carbohydrate-hydrolysing enzymes. The objective of this research was to evaluate the inhibitory effect of the hydroxytyrosol and the oleuropein against α-amylase and α-glucosidase. The hydroxytyrosol was purified from olive leaves. The result shows that the hydroxytyrosol had the strongest α-glucosidase inhibitory effect with IC50 values of 150 µM with mild inhibition against α-amylase. The enzyme kinetic studies, using Lineweaver-Burk indicated that, in the presence of the hydroxytyrosol, the Michaelis-Menton constant (Km) remained constant but the maximal velocity (Vmax) decreased, revealing a non-competitive type of inhibition with inhibition constants; Ki for the formation of the inhibitor-enzyme complex and Kis for the formation of the inhibitor-enzyme-substrate complex of 104.3 and 150.1 µM, respectively. On the other hand, oleuropein showedan uncompetitive inhibition. The concentrations used in this work were below cytotoxic levels observed at 400 µM. However, at 600 µM, the hydroxytyrosol significantly decreased viability of the Caco-2 cells (p < 0.05) and in the case of the oleuropein, there's an increase in cell number compared to control (p < 0.05). These results suggest that the hydroxytyrosol and oleuropein are two potential effective α-glucosidase inhibitors for management of postprandial hyperglycemia.

Valorization of the peel of pea: Pisum sativum by evaluation of its antioxidant and antimicrobial activities.

This study deals with evaluating antioxidant and antimicrobial activities of the peel of pea (Pisum sativum), with particular attention to the content of some bioactive compounds. Total content of polyphenols and flavonoids of Pisum sativum peel extracts, including a crude aqueous extract, a methanolic extract and an ethyl acetate extract was carried out according to the standard methods to assess their corresponding antioxidant activities. The organic solvents extracts antioxidant activities, determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay, ferric reducing (FRAP) assay and 2,2 azinobis 3-ethylbenzo-thiozoline-6-sulfonic acid (ABTS) assay, were relatively high. The highest activity was found in ethyl acetate extract. The antimicrobial activities of extracts were also assessed. The highest MIC value was occurred with E.Coli (850 µg/ml) when using ethyl acetate extract. From the results obtained, Pisum sativum peel can be considered as a very good source of health promoting compounds.

Antioxidant and lipase inhibitory activities and essential oil composition of pomegranate peel extracts.

The chemical composition of essential oil, antioxidant and pancreatic lipase inhibitory activities of various solvent extracts obtained from pomegranate peelTunisian cultivar was evaluated. Gas chromatography/mass spectrometry was used to determine the composition of the PP essential oil. Nine-teen components were identified and the main compounds were the camphor (60.32%) and the benzaldehyde (20.98%). The phenolic and flavonoids content varied from 0 to 290.10 mg Gallic acid equivalent and from 5.2 to 20.43 mg catechin equivalent/g dried extract. The antioxidant activity of various solvent extracts from pomegranate peel was also investigated using various in vitro assays as the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical method, ß-carotene bleaching and reducing power assays.Methanol and ethanol extracts showed the most potent antioxidant activity in all assays tested followed by water and acetone extracts. The inhibitory effect of the pomegranate peelextracts on porcine pancreatic lipase was evaluated and the results showed that ethanol and methanol extracts markedly reduced lipase activity. Generally, the highestlipase activity inhibitory (100%) was observed at a concentration of 1 mg/ml after 30 min of incubation. LC-MS analysis of ethanol extract showed the presence of four components which are cholorogenic acid, mannogalloylhexoside, gallic acid and ellagic acid. Our findings demonstrate that the ethanol extract from pomegranate peel might be a good candidate for furtherinvestigations of new bioactive substances.

Strategy for improving extracellular lipolytic activities by a novel thermotolerant Staphylococcus sp. strain.

BACKGROUND: Extracellular bacterial lipases received much attention for their substrate specificity and their ability to function under extreme environments (pH, temperature...). Many staphylococci produced lipases which were released into the culture medium. Reports of extracellular thermostable lipases from Staphylococcus sp. and active in alkaline conditions are not previously described. RESULTS: This study focused on novel strategies to increase extracellular lipolytic enzyme production by a novel Staphylococcus sp. strain ESW. The microorganism needed neutral or alkaline pH values between 7.0 and 12.0 for growth. For pH values outside this range, cell growth seemed to be significantly inhibited. Staphylococcus sp. culture was able to grow within a wide temperature range (from 30 to 55°C). The presence of oils in the culture medium leaded to improvements in cells growth and lipolytic enzyme activity. On the other hand, although chemical surfactants leaded to an almost complete inhibition of growth and lipolytic enzyme production, their addition along the culture could affect the location of the enzyme. In addition, our results showed that this novel Staphylococcus sp. strain produced biosurfactants simultaneously with lipolytic activity, when soapstock (The main co-product of the vegetable oil refining industry), was used as the sole carbon source. CONCLUSION: A simultaneous biosurfactant and extracellular lipolytic enzymes produced bacterial strain with potential application in soap stock treatment.

A newly high alkaline lipase: an ideal choice for application in detergent formulations.

BACKGROUND: Bacterial lipases received much attention for their substrate specificity and their ability to function in extreme environments (pH, temperature...). Many staphylococci produced lipases which were released into the culture medium. Reports of thermostable lipases from Staphylococcus sp. and active in alkaline conditions are not previously described. RESULTS: A newly soil-isolated Staphylococcus sp. strain ESW secretes an induced lipase in the culture medium. The effects of temperature, pH and various components in a detergent on the activity and stability of Staphylococcus sp. lipase (SL1) were studied in a preliminary evaluation for use in detergent formulation solutions. The enzyme was highly active over a wide range of pH from 9.0 to 13.0, with an optimum at pH 12.0. The relative activity at pH 13.0 was about 60% of that obtained at pH 12.0. It exhibited maximal activity at 60°C. This novel lipase, showed extreme stability towards non-ionic and anionic surfactants after pre-incubation for 1 h at 40°C, and relative stability towards oxidizing agents. Additionally, the crude enzyme showed excellent stability and compatibility with various commercial solid and liquid detergents. CONCLUSIONS: These properties added to the high activity in high alkaline pH make this novel lipase an ideal choice for application in detergent formulations.